|
Post by aqt on Apr 6, 2010 18:41:17 GMT -5
|
|
|
Post by aqt on May 11, 2010 14:52:50 GMT -5
Insect cells are a higher eukaryotic system than yeast and are able to carry out more complex post-translational modifications than the other two systems (see Comparison of Expression Systems). They also have the best machinery for the folding of mammalian proteins and, therefore, give you the best chance of obtaining soluble protein when you want to express a protein of mammalian origin. The most commonly used vector system for recombinant protein expression in insect is baculovirus, although baculoviral also can be used for gene transfer and expression in mammalian cells As with other eukaryotic expression systems, baculovirus expression of heterologous genes permits folding, post-translational modification and oligomerization in manners that are often identical to those that occur in mammalian cells. The insect cytoplasmic environment allows proper folding and S-S bond formation, unlike the reducing environment of the E. coli cytoplasm. Post-translational processing identical to that of mammalian cells has been reported for many proteins. These include proper proteolysis, N- and O-glycosylation, acylation, amidation, carboxymethylation, phosphorylation, and prenylation. Proteins may be secreted from cells or targeted to different subcellular locations. Single polypeptide, dimeric and trimeric proteins have been expressed in baculoviruses. Finally, expression of heterologous proteins is under the control of the strong polyhedrin promoter, allowing levels of expression of up to 30% of the total cell protein. www.genwaybio.com/gw_file.php?fid=6036 For example, inefficient secretion from insect cells may be circumvented by the addition of insect secretion signals (ex. honeybee melittin sequence). Improperly folded proteins and proteins that occur as intracellular aggregates may be due to expression late in the infection cycle. In such cases, harvesting cells at earlier times after infection may help. Low levels of expression can often be increased with optimization of time of expression and multiplicity of infection. The complete analysis of carbohydrate structures has been reported for a limited number of glycoproteins. Potential N-linked glycosylation sites are often either fully glycosylated or not glycosylated at all, as opposed to expression of various glycoforms that may occur in mammalian cells. Species-specific or tissue-specific modifications are unlikely to occur.
|
|
|
Post by aqt on May 11, 2010 14:58:05 GMT -5
tools.invitrogen.com/content/sfs/manuals/bevtest.pdfexpress heterologous genes in cultured insect cells and insect larvae. For large-scale applications, the baculovirus expression vector system by large scale applications do they mean global spraying via chemtrails? Oh, I'm sorry, I meant ILLEGAL AEROSOL OPERATIONS?aqt, mad as hell Hell hath no fury like a woman scorned? Hell hath no fury like a woman sickened by her government and brought back from near death with a vengence!!!!!!!!!!!
|
|
|
Post by aqt on May 11, 2010 17:16:51 GMT -5
|
|
|
Post by aqt on May 11, 2010 18:03:43 GMT -5
|
|
|
Post by aqt on May 11, 2010 18:16:02 GMT -5
fungus gnat larva looks very similar to out "tube within a tube" structures
|
|
|
Post by skyship on May 11, 2010 20:01:35 GMT -5
|
|
|
Post by aqt on May 12, 2010 11:35:18 GMT -5
Furthermore, we have shown that baculovirus can deliver foreign genes .... www.futuremedicine.com/doi/abs/10.2217/17460794.3.1.35?journalCode=fvlBaculovirus vector for gene delivery and vaccine development were genes delivered via baculovirus in an experimental vaccine? I contracted Morgellons shortly having Hepatitis B vaccine series...mandatory for my job position. aqt
|
|
|
Post by aqt on May 12, 2010 11:52:32 GMT -5
the fungus gnat is a relative of drosophelia melanogaster...fruit fly
in which we share a common (homeobox) gene.
aqt
|
|
|
Post by lilsissy on May 12, 2010 16:52:58 GMT -5
They also have the best machinery for the folding of mammalian proteins
Prions come to mind with that statement.
Jen
|
|
|
Post by skyship on May 12, 2010 20:56:57 GMT -5
Here is the MAIN one many are pushing: Arthrobotrys oligospora:
the spitzenkorper is at the tip. ============= check the names on this one who found this: www.youtube.com/watch?v=ikNGeJsvFu0&feature=relatedIts job is to destroy fungus: scialert.net/fulltext/?doi=ajps.2009.153.159&org=11=============== Now, we are at one edge of this bugger: thanks to Dr. Harvey, who found the pulmoni a while back, MRF, The c. pulmoni was also found in China, the black is male, female red, or vis a vis....... ================== Worm: c pulmoni, can be freeze dried, Fungus gnat, buffalo gnat, fungus bugs, and others carry and/or destroy the fungus, shore flies, there is one that has a stinger and can flip around, forgot the name Used on caterpillars. net: the fungus itself tip: spitzenkorper, in the fungus apical tip or core, of the creature. ========== Arthrobotrys oligospora:www.biological-research.com/Fungi/India__Nematodes_capture/india__nematodes_capture.htmlcaptures nematodes ========== could be in the trap itself: something to think about, did any of this come off our skin? ============= www.microbelibrary.org/ASMOnly/details.asp?id=483other things to think about. Fungus can kill ants, spiders, bugs, worms, however, seems the fungas gnats kill the fungus. =============== Genes from drosophila mel.. were models used to study molecular science. ============ compare drosophila melanogaster to fungus gnat: en.wikipedia.org/wiki/Drosophila_melanogasterfr.treknature.com/gallery/Europe/Switzerland/photo87571.htmfungus gnat larvae: note size: culturesheet.org/pests:fungus_gnats============ Homeobox: Model organisms used to study humans and genes other organisms have that humans have. en.wikipedia.org/wiki/Category:Model_organism_databasesnext category Molecular cells. ============ Could the eggs be in the fungus itself? not the gnat? seems to be kicking them out. parasitoid. The gnat would be the destroyer of the fungus, and the fungus the destroyer of the worm. ============== Shore flies are not fungus gnats! I am a reviewer and critic at this point: suggestive, supportive, objective criticism, needed for pure truth here! I am one of those old writers, who reveiws everything! Do not mean to offend anyone, but, a perspective is needed. floriculture.osu.edu/archive/dec96/gnats.htmlThe videos speak for themselves. Thankyou All for helping us see all of this puzzle. Conclusions can only be reached when a hypothesis can be presented and proven. Without a hypothesis, the videos, photos and the like are pure observations, which are the best EVIDENCE their is, but, has to be attached to human somehow. skyship
|
|
|
Post by skyship on May 12, 2010 21:52:00 GMT -5
OLIGO: Polyuridylation, also called oligouridylation, is the addition of several uridine nucleotides to the 3' end of an RNA. ... en.wikipedia.org/wiki/Oligo(U)_tail www.google.com/search?hl=en&q=define%3A+oligo&aq=0&aqi=l1g1&aql=&oq=oligo+definition&gs_rfai=------------------------ polyuridylation: Polyuridylation, also called oligouridylation, is the addition of several uridine nucleotides to the 3' end of an RNA. One group of RNAs that can be polyuridylated are histone mRNAs that lack a poly(A) tail. Polyuridylation of a histone mRNA promotes its degradation, involving the exosome. Other RNAs in Arabidopsis and mouse have been seen to be polyuridinylated after cleavage.[1] ================== New ways to meet your (3′) end—oligouridylation as a step on the path to destruction
1. Carol J. Wilusz2 and 2. Jeffrey Wilusz1 - Author Affiliations 1. Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523, USA Messenger RNA degradation is a vital contributor to the control of gene expression that generally involves removal of a poly(A) tail in both prokaryotes and eukaryotes. In a thought-provoking study in this issue of Genes & Development, Mullen and Marzluff (2008) present data supporting a novel mechanism of mRNA decay. They discovered that histone mRNAs, which are unique in that they are never polyadenylated in mammalian cells, degrade by a cell cycle-regulated mechanism that involves addition of a short oligo(U) tail at the 3′ end. Interestingly, this oligo(U) tract is recognized by the Lsm1–7 complex, which then appears to feed the transcript into the standard mRNA decay pathways. These findings are exciting because they invoke parallels with prokaryotic mRNA decay, which requires polyadenylation immediately prior to degradation and involves an Lsm homolog, Hfq. Moreover, recent studies have identified other oligouridylated RNAs and several poly(U) polymerases, implying that this may be a more widespread mechanism for turnover of RNA. genesdev.cshlp.org/content/22/1/1.long======================= Uridine nucleotides:uridine /uri·dine/ (ūr´ĭ-dēn) a pyrimidine nucleoside containing uracil and ribose; it is a component of nucleic acid and its nucleosides are involved in the biosynthesis of polysaccharides. Symbol U. uridine diphosphate (UDP) a pyrophosphate-containing nucleotide that serves as a carrier for hexoses, hexosamines, and hexuronic acids in the synthesis of glycogen, glycoproteins, and glycosaminoglycans. uridine monophosphate (UMP) uridylic acid; a nucleotide, uridine 5′-phosphate. uridine triphosphate (UTP) a nucleotide involved in RNA synthesis.Dorland's Medical Dictionary for Health Consumers. © 2007 by Saunders, an imprint of Elsevier, Inc. All rights reserved. u·ri·dine (yr-dn) n. A white odorless powder that is the nucleoside of uracil and is important in carbohydrate metabolism. medical-dictionary.thefreedictionary.com/uridine=========== OligomerIn chemistry, an oligomer consists of a few monomer units (ολιγος, or oligos, is Greek for "a few"), in contrast to a polymer that, at least in principle, consists of an unlimited number of monomers.[1] Dimers, trimers and tetramers are oligomers. Many oils are oligomeric, such as liquid paraffin. Plasticizers are oligomeric esters widely used to soften thermoplastics such as PVC. They may be made from monomers by linking them together, or by separation from the higher fractions of crude oil. Polybutene is an oligomeric oil used to make putty. Greek prefixes are often used to designate the number of monomer units in the oligomer, for example a tetramer being composed of four units and a hexamer of six. In biochemistry, the term oligomer is used for short, single-stranded nucleic acid fragments, such as DNA or RNA, or similar fragments of analogs of nucleic acids such as peptide nucleic acid or Morpholinos. Such oligos are used in hybridization experiments (bound to glass slides or nylon membranes), as probes for in situ hybridization or in antisense experiments such as gene knockdowns. It can also refer to a protein complex made of two or more subunits. In this case, a complex made of several different protein subunits is called a hetero-oligomer or heteromer. When only one type of protein subunit is used in the complex, it is called a homo-oligomer or homomer. Oligomerization is a chemical process that converts monomers to a finite degree of polymerization. The actual figure is a matter of debate, often a value between 10 and 100. When an oligomer forms as a result of chain transfer the oligomer is called a telomer and the process telomerization.[2] A telomere is a region of highly repetitive DNA at the end of a linear chromosome. en.wikipedia.org/wiki/Oligomer================= fungus are us!.............flies are us...........worms are us............plants are us........... roots are us, roots of ganglion are what? diatoms are us......algae are us.... thermophiles are us........iron is us..........copper is us ..........aluminum is us.......... zinc is us..............electrons are us............protons are us. uridine is us........ radiative particles are us.............................................................................. Homeobox genes, homeobox chemistry, homeobox radiative particles........
The spark of life, the filament of life, mycelia or rhizobias................
at some point you will see.
============= oligos are artificial genes......=============== mixtures of natural and artificial:
Many of the enabling technologies for synthetic biology have existed for several years. The metabolic engineering of bacteria for natural product synthesis was first achieved in the early 1970s, and engineered bacterial plasmids for biotechnology were developed during the 1980s. Genetically modified organisms with relatively sophisticated systems for gene expression and containment have been around for nearly as long. The main difference between genetic engineering and synthetic biology is that whereas the former involves the transfer of individual genes from one species to another, the latter envisions the assembly of novel microbial genomes from a set of standardized genetic parts. These components may be natural genes that are being applied for a new purpose, natural genes that have been redesigned to function more efficiently, or artificial genes that have been designed and synthesized from scratch. Synthetic genomics” refers to the set of technologies that makes it possible to construct any specified gene (or full genome) from short strands of synthetic DNA called “oligonucleotides,” which are produced chemically and are generally between 50 and 100 base-pairs in length. In August 2002, Eckard Wimmer, a virologist at the State University of New York at Stonybrook, announced that over a period of several months his research team had assembled live, infectious poliovirus from customized oligonucleotides mail-ordered from a commercial supplier, using a map of the viral genome available on the Internet [13]. In 2003, Hamilton Smith and his colleagues at the Venter Institute developed a faster method for genome assembly, using synthetic oligonucleotides to construct a bacteriophage called φX174 (containing 5,386 DNA base-pairs) in only two weeks [14]. Most recently, in 2005, scientists at the U.S. Centers for Disease Control and Prevention synthesized the so-called Spanish influenza virus which was responsible for the 1918-19 flu pandemic that killed between 50 million and 100 million people worldwide [15, 16]. www.thenewatlantis.com/publications/the-promise-and-perils-of-synthetic-biologyskyship
|
|
|
Post by skyship on May 12, 2010 21:53:39 GMT -5
|
|
|
Post by skyship on May 12, 2010 22:35:32 GMT -5
|
|
|
Post by skyship on May 13, 2010 16:07:57 GMT -5
According to this baculovirus, all 5 lines, are not pathagenic to verterbrates and plants and they do not employ transformed cells. pubs.acs.org/doi/abs/10.1021/bp00027a012============ So, if this is true, then we must look at the homologenous genes used, the ones that from ancient times have been consistent with man. Yeast, S. cerev is one, so if we find the homeobox again, we can see what proteins were used for what in humans, that are supposed to be conserved in man. Lets list those, and find the homologenous genes, those are the only ones that can mimic ours. Baculo is used in labs. However, its used in vaccines is one we should be concerned about. How can you maim a microbe or parasite with a vaccine? It will not kill it, it will maim or change it, the dying parasite like Oncho, can cause calcification, microbes the same. However, if poly A or dimers are formed, they are formed from the protein or DNA, so somehow that foreign DNA is getting into us. The parasite is what needs to be looked at, and this has been ignored. Microbial parasite, that self assembles, from a core. Apical core of life? delta F508 in CFTR ΔF508 (delta-F508) is a specific mutation within the human genome. The mutation is a deletion of three base pairs at position 508 in the nucleotide sequence of a protein called the cystic fibrosis transmembrane conductance regulator (CFTR). The deletion prevents the codon for phenylalanine (F) from obtaining its normal position. Having two copies of this mutation, inherited from both parents, is the leading cause of cystic fibrosis (CF) Mechanism The three DNA base pairs T-A-G at position 507 of the CFTR nucleotide sequence form the template for the mRNA codon A-U-C for the amino acid isoleucine, while the three DNA base pairs A-A-A at the adjacent position 508 form the template for the codon U-U-U for phenylalanine. The ΔF508 mutation is a deletion of the G pair from position 507 along with two A-A pairs from position 508, leaving the DNA sequence T-A-A at position 507 forming the codon A-U-U. Since A-U-U also codes for isoleucine, position 507's amino acid is unchanged, and the mutation's net effect is equivalent to a deletion ("Δ") of the sequence resulting in the codon for phenylalanine ("F") at position 508. Prevalence ΔF508 is present in approximately one in 30 Caucasians. Scientists have estimated that the mutation occurred over 52,000 years ago in Northern Europe. From an evolutionary standpoint the mutation's negative effects (see below) are outweighed by the fact that it reduces water-loss during cholera, a common cause of death in Europe when the mutation first appeared. Effects The CFTR protein—when in the proper position—opens channels in the cell wall which release chloride ions out of the cells. This causes osmosis to draw water out of the cell. The ΔF508 mutation can prevent the CFTR from moving into its proper position in the cell. en.wikipedia.org/wiki/%CE%94F508===================== An old diseases, and not studied much is Panniculosis: Chronic stages of the disease typically coincide with the formation of a structure known as a pannus. A pannus is a membrane of granulation tissue composed of mesenchyme- and bone marrow-derived cells. Formation of the pannus stimulates the release of IL-1, platelet-derived growth factor, prostaglandins, and substance P by macrophages, which ultimately cause cartilage destruction and bone erosion. en.wikipedia.org/wiki/Pannus===================== this traces back to hsp70 and others, the element in the heat shock proteins??? DnaK, DnaJ involved. Where did the core for heat shock proteins come in? Where did this heat shock protein come from? why is it everywhere? hsp10: mips.helmholtz-muenchen.de/genre/proj/yeast/searchEntryAction.do?text=YOR020cnote homologues in basidiomyctes and others. skyship
|
|
|
Post by kammy on May 19, 2010 14:07:23 GMT -5
Hi guys/gals - I'm glad to see you started this thread, I'm still around - just busy these days. I'm trying to see if I can repeat last year's experiments where the petri dishes turned up with live insects. This year I'm keeping the dishes taped shut, inside a plastic container - I am waiting on temperature outside to reach the 90's and the end of July timeframe to see if this happens again. I'll keep you posted.
|
|
|
Post by skyship on May 19, 2010 23:38:02 GMT -5
And that my friend, may be the key, the temperature.
I bet, it has a lot to do with how this organism unfolds.
Seems they can build an insect, like a weird, wormy fungal microbe.
Good to see you again.
sounds like a grand experiment.
Skyship
|
|
|
Post by aqt on May 28, 2010 18:41:17 GMT -5
This insect "crawled" out of a Morgellons sufferer's eye
|
|
|
Post by kammy on May 28, 2010 20:39:13 GMT -5
And that my friend, may be the key, the temperature. I bet, it has a lot to do with how this organism unfolds. Seems they can build an insect, like a weird, wormy fungal microbe. Good to see you again. sounds like a grand experiment. Skyship We need some Americans to help with the experiment. We have Europe conducting it, Jo in England and myself in Germany. Someone needs to purchase a few petri dishes soon... ??
|
|
|
Post by skyship on May 31, 2010 20:28:24 GMT -5
The human baculovirus evidently is harbored in this: Epstein Barr virus episome.================ Baculoviruses are a large group of insect infecting DNA viruses characterized by the rod‐shaped morphology of their virion. They have been used as biopesticides and as tools for efficient recombinant protein production in insect cells . Baculoviruses are also capable of entering into a wide spectrum of vertebrate cells making them a potential new tool for gene delivery.In this work, we developed novel baculoviral vectors for enhanced and longterm gene delivery. We studied the feasibility of post‐transcriptional regulatory element and optimized culture conditions in order to enhance transgene expression of baculovirus. The mechanism behind the positive medium effect is probably due to the reorganization of intermediate filaments (vimentin), which leads to improved access of baculoviral capsids into the nucleus. In addition, we created a baculovirus‐Epstein‐Barr hybrid virus which combines the advantages of the baculovirus vector and prolonged episomal gene expression achieved by the Epstein‐Barr virus‐based episome.
pages 20-21 epublications.uef.fi/pub/urn_isbn_978-952-61-0022-7/urn_isbn_978-952-61-0022-7.pdf================== What is this made of?
the Epstein‐Barr virus‐based episome. They say this is used for therapy, like repairing dna mutations, but, if everyone has this this forms a base for something to be activated again. Along with Epstein barr, Cytomegalo virus was used as well.
======= Cytomegalo virus in baculovirus, clathrin coated ap1 and ap3
Immune system does not recognize these viruses: "Viral Subversion of the Immune SystemDomenico Tortorella, Benjamin E. Gewurz, Margo H. Furman, Danny J. Schust, and Hidde L. Ploegh Department of Pathology, Harvard Medical School, Boston, Massachusetts, 02115; email: dtortorella@hms.harvard.edu, bgewurz@student.med.harvard.edu, mfurman@fas.harvard.edu, dschust@hms.harvard.edu, hploegh@hms.harvard.edu This review describes the diverse array of pathways and molecular targets that are used by viruses to elude immune detection and destruction. These include targeting of pathways for major histocompatibility complex-restricted antigen presentation, apoptosis, cytokine-mediated signaling, and humoral immune responses. The continuous interactions between host and pathogens during their coevolution have shaped the immune system, but also the counter measures used by pathogens. Further study of their interactions should improve our ability to manipulate and exploit the various pathogens." ============= So, while the pathogens are there they are used to manipulate and introduce the second messenger, mRNa.==================== ======================== The binding of AP-1 clathrin adaptor particles to Golgi membranes requires ADP-ribosylation factor, a small GTP-binding protein
Mark A. Stamnes and James E. Rothman Program in Cellular Biochemistry and Biophysics Rockefeller Research Laboratory Sloan-Kettering Institute Memorial Sloan-Kettering Cancer Center New York, New York 10021 USA * Abstract * The small GTP-binding protein, ADP-ribosylation factor (ARF), has previously been shown to mediate the binding to Golgi membranes of the coatomer of nonclathrin-coated (COP-coated) vesicles. We now report that ARF is also required for the binding of the AP-1 adaptor protein of clathrin-coated vesicles to Golgi membranes. The binding of coat proteins from both clathrin- and COP-coated vesicles requires an additional Golgi membrane-associated factor. These results suggest that a mechanistic similarity underlies diverse types of vesicle coats.www.cell.com/retrieve/pii/009286749390277W================ ============ [b ] adaptor protein[/b] An adaptor protein links components in signaling pathways by acting as an accessory to the chief proteins in a signal transduction pathway.▼ adaptor genes : adaptor proteins : AP-1, AP-2 : Cbl : clathrin : Grb2 : LIME : Nck : RING : SH2, SH3 : thymocyte development : TIR : TLR : Toll-like receptor : ubiquitin-ligases▼ Adaptor proteins lack intrinsic enzymatic activity, instead they mediate specific protein-protein interactions that drive the formation of protein complexes. The specificity of Toll-like receptor signaling occurs by means of adaptor proteins possessing Toll–interleukin 1 receptor (TIR) domains. Examples of adaptor proteins include:Adaptor protein complex 1 ( AP-1) and AP-2, which interact with clathrin-coated vesicles located at the trans-Golgi network (TGN) or the plasma membrane. Heterotetramer adaptor protein complexes that function as vesicle coat components and cargo molecule selectors in various membrane trafficking pathways. Cbl adaptor proteins are RING-type E3 ubiquitin ligases that may be involved in the negative regulation of thymocyte development, targeting its substrate for ubiquitination [ 1 ]. The ubiquitin ligase activity of Cbl, and of its homologue Cbl-b, plays a role in the negative regulation of upstream kinases, such as Lck, Syk and PI3K, in T and B cellsGrb2IPS-1/MAVS/VISA/Cardif LIME (Lck-interacting molecule) is a raft-associated adaptor protein that is expressed predominantly in T lymphocytes, and which becomes tyrosine phosphorylated after cross-linking of the CD4 or CD8 coreceptors. Phospho-LIME associates with the Src family kinase Lck and its negative regulator, Csk. NckSrc homology = Shc proteins → SH2 recognize phosphotyrosine residues and SH3 domains recognise proline-rich sequences Genes encoding adaptor proteins include:GRAP → GRB2-related adaptor protein GRAP2 → GRB2-related adaptor protein 2 LDLRAP1 → low density lipoprotein receptor adaptor protein 1 NCK1 → NCK adaptor protein 1 NCK2 → NCK adaptor protein 2 NOS1AP → nitric oxide synthase 1 (neuronal) adaptor protein PIK3AP1 → phosphoinositide-3-kinase adaptor protein 1 SH2B1 → SH2B adaptor protein 1 SH2B2 → SH2B adaptor protein 2 SH2B3 → SH2B adaptor protein 3 SHB → Src homology 2 domain containing adaptor protein B SLC4A1AP → solute carrier family 4 (anion exchanger), member 1, adaptor protein tags [Proteins] [adaptor+protein] Labels: adaptor protein, Cbl, clathrin-coated vesicles, Grb2, IL, Nck, TGN, TLR proteian.blogspot.com/2007/12/adaptor-protein.html#adaptor-proteins-------------------- Getting complicated, but can see the construction from the Triskelion which forms the shhhhhhhhhh.. BB..........
============= What does this form?www.youtube.com/watch?v=eRslV6lrVxY&feature=related================== back later, more deciphering..........
skyship
|
|
|
Post by skyship on May 31, 2010 23:23:51 GMT -5
baculovirus‐Epstein‐Barr hybrid virus Since we already have Epstein Barr, is is utilized with baculovirus. It even has a name: GP350 ============== Here it is used in a vaccine for mononucleiosis: www.ncbi.nlm.nih.gov/pubmed/18190254---------------------- What else is it used for? ---------- These data thus suggest that HABP-11382, -11389, and -11416 sequences are involved in EBV binding and invasion of B-cell lymphocytes and could be used not only for designing strategies against EBV-induced diseases (i.e. EBV-induced post-transplant lymphoproliferative disorder), but also for designing prophylactic EBV vaccine candidates. www.jbc.org/content/280/42/35598.full======================= many of the uses of this have to do with transplants, and vaccines. ================= That would be a way into the human without bug vectors, just using the bucky ball. ================ skyship
|
|
|
Post by skyship on Jun 1, 2010 0:23:42 GMT -5
A Geometric Principle May Guide Self-Assembly of Fullerene Cages from Clathrin Triskelia and from Carbon Atoms" Clathrin triskelia and carbon atoms alike self-assemble into a limited selection of fullerene cages (with n three connected vertices, 3n/2 edges, 12 pentagonal faces, and (n−20)/2 hexagonal faces). We show that a geometric constraint—exclusion of head-to-tail dihedral angle discrepancies (DADs)—explains this limited selection as well as successful assembly into such closed cages in the first place. An edge running from a pentagon to a hexagon has a DAD, since the dihedral angles about the edge broaden from its pentagon (tail) end to its hexagon (head) end. Of the 21 configurations of a central face and surrounding faces, six have such DAD vectors arranged head-to-tail. Of the 5770 mathematically possible fullerene cages for n ≤ 60, excluding those with any of the six configurations leaves just 15 cages plus buckminsterfullerene (n = 60), among them the known clathrin cages. Of the 216,739 mathematically possible cages for 60 < n ≤ 84, just the 50 that obey the isolated-pentagon rule, among them known carbon cages, pass. The absence of likely fullerenes for some n (30,34,46,48,52–58,62–68) explains the abundance of certain cages, including buckminsterfullerene. These principles also suggest a “probable roads” path to self-assembly in place of pentagon-road and fullerene-road hypotheses."www.ncbi.nlm.nih.gov/pmc/articles/PMC2186234/More..... ========= ...."Although the term “fullerene” was applied initially by chemists to closed cages assembled from carbon atoms in 1985 (1), biologists were already familiar with fullerene cages (2,3) assembled from the protein clathrin (4,5). Such cages have n three-connected vertices, 3/2n edges, 12 pentagonal faces, and (n − 20)/2 hexagonal faces. Moreover, both carbon atoms, each just 12 Daltons, and clathrin trimers (triskelia) (6–8), each with mass ∼50,000 times as much (6–11), self-assemble into fullerene cages of a variety of sizes and shapes (2–5,12,13). Carbon does this from the gas phase at temperatures around 1200°C (14–16). Clathrin does this inside cells, where it self-assembles around inwardly budding patches of membrane (13,17) to encapsulate vesicles of different sizes (2,3,5,18,19)" ..."It does so at two sites: It withdraws such patches from successive compartments of the trans-Golgi network and from the plasma membrane (5,13,20,21). At the latter site, the area of synaptic vesicle membrane taken up by clathrin-mediated endocytosis determines the size of neurotransmitter-filled synaptic vesicles and the amount of neurotransmitter in each quantum that is subsequently released at the synapse ".....------- ......required for endocytosis.............
------- ..."Carbon famously self-assembles into this C60 cage, which the chemists named “buckminsterfullerene” or the “buckyball” (1). Carbon also self-assembles into many larger cages, including the one IPR isomer of C70 (27), very large, elongated nanotubes (28),
and the small 36-15 cage (29,30). Clathrin also self-assembles into nanotube-type structures (31)."...... ---------- ..."Self-assembly is the very basis for life. Small multisubunit proteins, the shell of a virus particle, the complex molecular factory that is the ribosome, and even large structures like organelles can be regarded as products of self-assembly; so, for that matter, can the folding of an individual protein molecule. The usual outcome in these examples is a single structure. By contrast, both carbon atoms and clathrin triskelia self-assemble into a variety of sizes and shapes of (fullerene) structures, so the many completed, probable cages at the ends of probable roads must occupy local energy minima rather than a global energy minimum. Thus, probable roadmaps—actually potential energy landscapes (78) that combine kinetics and equilibrium thermodynamics to guide self-assembly and equilibrium thermodynamics to drive it—may be able to describe a broad range of self-assembly processes." ===================== So, which is which? Are carbon buckyballs substituting for clathrin coated vesicles? Or are they making them? or are the clathrin coated vesicles the buckyball forming itself?
==================== triskeliari·skel·i·on (tr-skl-n, tr-) also tri·skele (trskl, trskl) n. pl. tri·skel·i·a (-skl-) also tri·skeles A figure consisting of three curved lines or branches, or three stylized human arms or legs, radiating from a common center.[New Latin, from Greek triskels, three-legged : tri-, tri- + skelos, leg.] The American Heritage® Dictionary of the English Language, Fourth Edition copyright ©2000 by Houghton Mifflin Company. Updated in 2009. Published by Houghton Mifflin Company. All rights reserved. triskelion [trɪˈskɛlɪˌɒn -ən], triskele [ˈtrɪskiːl] n pl triskelia [trɪˈskɛlɪə], triskeles (Fine Arts & Visual Arts / Art Terms) a symbol consisting of three bent limbs or lines radiating from a centre[from Greek triskelēs three-legged, from tri- + skelos leg] Collins English Dictionary – Complete and Unabridged © HarperCollins Publishers 1991, 1994, 1998, 2000, 2003 ThesaurusLegend: Synonyms Related Words Antonyms Noun 1. triskelion - a figure consisting of three stylized human arms or legs (or three bent lines) radiating from a centertriskele pattern, design, figure - a decorative or artistic work; "the coach had a design on the doors"www.thefreedictionary.com/triskelia=============== Like the buckyball has design on the human================= Skyship
|
|
|
Post by skyship on Jun 1, 2010 1:39:55 GMT -5
If fullerene nanoparticles are at the core of clathrin coated triskelions, and carbon bucky balls, then? ??....... ========= The translocation of fullerenic nanoparticles into lysosome via the pathway of clathrin-mediated endocytosis "Manufactured fullerene nanoparticles easily enter into cells and hence have been rapidly developed for biomedical uses. However, it is generally unknown which route the nanoparticles undergo when crossing cell membranes and where they localize to the intracellular compartments. Herein we have used both microscopic imaging and biological techniques to explore the processes of [C60(C(COOH)2)2]n nanoparticles across cellular membranes and their intracellular translocation in 3T3 L1 and RH-35 living cells. The fullerene nanoparticles are quickly internalized by the cells and then routed to the cytoplasm with punctate localization. Upon entering the cell, they are synchronized to lysosome-like vesicles. The [C60(C(COOH)2)2]n nanoparticles entering cells are mainly via endocytosis with time-, temperature- and energy-dependent manners. The cellular uptake of [C60(C(COOH)2)2]n nanoparticles was found to be clathrin-mediated but not caveolae-mediated endocytosis. The endocytosis mechanism and the subcellular target location provide key information for the better understanding and predicting of the biomedical function of fullerene nanoparticles inside cells." iopscience.iop.org/0957-4484/19/14/145102;jsessionid=B259F0FE6A0EFA64D0D9BD39D4F9FF10.c1skyship
|
|
|
Post by Southcty on Sept 29, 2010 17:24:05 GMT -5
That looks like a bedbug. This insect "crawled" out of a Morgellons sufferer's eye
|
|
|
Post by skyship on Oct 1, 2010 1:14:38 GMT -5
Baculoviruse used was P35 Baculovirus P35 interacts with a subunit of human RNA polymerase II and can enhance promoter activity in human cells The early protein P35 from the baculovirus Autographa californica nucleopolyhedrovirus is a direct inhibitor of caspases and can block apoptosis in a wide variety of systems.,,,,,,' ,,,,, identified the RPB11a subunit of human RNA polymerase II as an interaction partner of P35. Specificity of the interaction was confirmed by affinity blotting. By immunocytology, P35 was in part found in the nucleus of transfected cells. Homology searches further revealed that P35 has structural similarity with RPB3, the subunit of RNA polymerase II that has been demonstrated to interact directly with RPB11a. When transfected into human colon carcinoma cells, P35 was able to enhance the activity of E-cadherin and {beta}-actin promoters by about a factor of two as measured by luciferase reporter assay. P35 and hRPB11a together enhanced the E-cadherin activity about three- to fourfold. These data suggest an additional role for P35 in the regulation of cellular transcription. vir.sgmjournals.org/cgi/content/full/84/11/3011 protein P35 from the baculovirus Autographa californica nucleopolyhedrovirus
|
|
|
Post by skyship on Oct 1, 2010 15:20:05 GMT -5
Our report of sup46 Het-s causes prions to form and amyloid fibers
Now the P35 from aplysia used in baculovirus for humans.
Now the ferritin and I know there was an operation that put iron particles in ocean to clean ship wrecks, so no one could find some truth articles in some of those wrecks.
No, was to clean up the environment, they say.===================== Electrokinetic Remediation of Cadmium-Contaminated Soil Using Zero-Valent Iron Particles ABSTRACT Electrokinetic remediation technique (Ek) has shown impressive results in soil remediation. Several research and experimental work have been carried out recently to enhance the Electrokinetic remediation process. There are certain enhancing techniques which have been developed like pH control, desorbing agent concentration, washings solution and use of zero valent iron particle as a permeable reactive barrier. Due to their minute size and higher surface area zero valent iron particle become highly reactive in EkR process. The use of iron particle as a permeable reactive barrier as shown remarkable result in enhancing the Electrokinetic remediation process. The present paper deals with the use of zero-valent iron (ZVI) particle as a permeable reactive barrier and its effect on Ek process for cadmium contaminated soil. Three experiments have been conducted: first the soil is washed with distilled water with distilled water used as a purging solution; second the soil is washed with distilled water with ammonium citrate used as purging solution; third the soil is washed with distilled water with ammonium citrate used as purging solution and a permeable reactive barrier of zero valent iron particle has been created. The results show that without the use of zero valent iron powder 62% removal efficiency has been recorded and with use ZVI as permeable reactive barrier 83% removal efficiency has been recorded. The present work is a progressive research work at MNNIT Allahabad India, on the subject of Electrokinetic remediation of fine grained soil now that the Electrokinetic process has been enhanced by using ZVI. KEYWORDS: Ammonium citrate, cadmium, Electrokinetic remediation, ZVI. -- "In the human body, cadmium accumulates mainly in the kidneys. At high levels, it can reach a critical threshold and can lead to serious kidney failure. Recent studies (Buchet & Bernard 1998) have shown that kidney effects may be reversible at low exposures once cadmium exposure is reduced or removed. "www.ejge.com/2010/Ppr10.015/Ppr10.015.pdf=========== If you remember CeSe Zn0s were used to clean up the environment, then along came electrical cleaning up of the environment, by using iron particles, provides an electrical magnetic component.
But this was to clean up cadmium, how did it get there in the first place?
From the previous aerosol operations, using, CeSe ZnO buckyballs. Tellers idea.
Excessive selenium causes pellagra.......... and cadmium?
well, so the Ce was used to clean up the Klebsiella aerogenes.
Aerogenes, again from some aerosol operation plan.......
so, we just have to follow the trail. You can see where it is eating through cement, roads, etc. the brown spots on the cement?
================ this was 1983.................. Inorganic Phosphate Accumulation and Cadmium Detoxification in Klebsiella aerogenes NCTC 418 Growing in Continuous CultureKlebsiella aerogenes NCTC 418, growing in the presence of cadmium under glucose-, sulfate-, or phosphate-limited conditions in continuous culture, exhibits two different cadmium detoxifying mechanisms. In addition to sulfide formation, increased accumulation of Pi is demonstrated as a novel mechanism. Intracellular cadmium is always quantitatively counterbalanced by a concerted increase in both inorganic sulfide and Pi contents of the cells. This led to the conclusion that production of sulfide and accumulation of Pi are detoxification mechanisms present in K. aerogenes but that their relative importance is crucially dependent on the strain and the growth conditions employed. www.ncbi.nlm.nih.gov/pmc/articles/PMC239677/pdf/aem00159-0160.pdf==================== micron sub particle ? accumulation of Pi =========== tinyurl.com/24omkts"Accumulation of Pi-3 phosphates in plasma membranes leads to activation of several kinases"......... Protein kinase B(PKB) these become localized in the "plasma membrane" ============== so the sub micron particle is related to phosphates as well as cadmium iron etc. but, it is these Klebsiella aerogenes: ========= Black specks are the particles........ ============ remember the cryptococcus gatti, a derivative of this Cryptococcus neoformans. ================ Induction by Klebsiella aerogenes of a Melanin-Like Pigment in Cryptococcus neoformansThis is why our melanin is being deleted!
============ While studying the interaction of Cryptococcus neoformans with Dictyostelium discoideum, we noticed that yeast colonies in agar with a feeder lawn of Klebsiella aerogenes were brown. This finding was intriguing because C. neoformans colonies are not pigmented unless they are provided with precursors for melanization. Strains of all C. neoformans serotypes produced brown pigment in response to K. aerogenes at 22, 30, and 37°C. Pigment production required fungal laccase and was suppressed by high concentrations of glucose. Treatment of brown cells with guanidinium isothiocyanate and hot concentrated HCl yielded particulate material that had the physical and chemical characteristics of melanins. No pigment formation was observed when C. neoformans was exposed to live Escherichia coli or heat-killed K. aerogenes. Analysis of K. aerogenes supernatants revealed the presence of dopamine, which can be a substrate for melanin synthesis by C. neoformans. Our findings illustrate a remarkable interaction between a pathogenic fungus and a gram-negative bacterium, in which the bacterium produces a substrate that promotes fungal melanization. This observation provides a precedent that could explain the source of a substrate for C. neoformans melanization in the environment.aem.asm.org/cgi/content/full/72/2/1542skyship
|
|
|
Post by skyship on Oct 1, 2010 15:52:03 GMT -5
P35, CeSe ZnO, sup 35, aerogenes, ZVI Introduction of Bacteriophage Mu into Bacteria of Various Genera and Intergeneric Gene Transfer by RP4: :Mu YOSHIKATSU MUROOKA,* NOBORU TAKIZAWA, AND TOKUYA HARADA The Institute of Scientific and Industrial Research, Osaka University, Suita, Osaka (565), Japan The host range of coliphage Mu was greatly expanded to various genera of gram-negative bacteria by using the hybrid plasmid RP4::Mu cts, which is temperature sensitive and which confers resistance to ampicillin, kanamycin, and tetracycline. These drug resistance genes were transferred from Escherichia coli to members of the genera Klebsiella, Enterobacter, Citrobacter, Salmonella, Proteus, Erwinia, Serratia, Alcaligenes, Agrobacterium, Rhizobium, Pseudo- monas, Acetobacter, and Bacillus. Mu phage was produced by thermal induction from the lysogens of all these drug-resistant bacteria except Bacillus. Mu phage and RP4 or the RP4: :Mu plasmid were used to create intergeneric recombinant strains by transfer of some genes, including the arylsulfatase gene, between Klebsiella aerogenes and E. coli. Thus, genetic analysis and intergeneric gene transfer are possible in these RP4: :Mu-sensitive bacteria. jb.asm.org/cgi/reprint/145/1/358.pdfIf this is in insects, and the baculo was used for construction of this, it is very hard to locate the causes, because of the polymorphisms that happen. skyship
|
|
|
Post by skyship on Oct 1, 2010 16:32:56 GMT -5
These drug resistance genes were transferred from Escherichia coli to members of the genera Klebsiella, Enterobacter, Citrobacter, Salmonella, Proteus, Erwinia, Serratia, Alcaligenes, Agrobacterium, Rhizobium, Pseudo- monas, Acetobacter, and Bacillus. Mu phage was produced by thermal induction from the lysogens of all these drug-resistant bacteria except Bacillus. Mu phage and RP4 or the RP4: :Mu plasmid were used to create intergeneric recombinant Mu phage was produced by thermal induction from the lysogens en.wikipedia.org/wiki/Lambda_phage====================== lysogens: comes back to the prophages: the baculo is a virus can be put in a phage Llambda phage: MU phages etc. the mutator, used to change genes, but must have a prion to unfold and fold the proteins. that is in the sup 35, the het-s creates the prions. Not the normal CJF but the one attached to the mutator. the vectors: ===================== Rapid Construction of Adenoviral Vectors by Lambda Phage GeneticsContinued improvements of adenoviral vectors require the investigation of novel genome configurations. Since adenovirus can be generated directly by transfecting packaging cell lines with viral genomes isolated from plasmid DNA, it is possible to separate genome construction from virus production. In this way failure to generate a virus is not associated with an inability to generate the desired genome. We have developed a novel lambda-based system that allows rapid modification of the viral genome by double homologous recombination in Escherichia coli. The recombination reaction and newly generated genome may reside in a recombination-deficient bacterial host for enhanced plasmid stabilityThe strategy relies on four main steps: (i) homologous recombination between an adenovirus cosmid and a donor plasmid (the donor plasmid carries the desired modification and flanking regions of homology to direct its recombination into the viral genome);
(ii) in vivo packaging of the recombinant adenoviral cosmids during a productive lambda infection;
(iii) transducing a recombination-deficient E. coli lambda lysogen with the generated lysate (the lysogen inhibits the helper phage used to package the recombinant andenoviral cosmid from productively infecting and destroying the host bacteria);
(iv) effectively selecting for the desired double-recombinant cosmid. Approximately 10,000 double-recombinant cosmids are recovered per reaction with essentially all of them being the correct double-recombinant molecule.
This system was used to generate quickly and efficiently adenoviral genomes deficient in the E1/E3 and E1/E3/E4 regions. The basis of this technology allows any region of the viral genome to be readily modified for investigation of novel configurations.
www.ncbi.nlm.nih.gov/pmc/articles/PMC136085/
=============
skyship
|
|
|
Post by skyship on Oct 1, 2010 16:50:00 GMT -5
Am trying to find link between use of baculovirus for specific link to lysogen, means lyses,
starting at the beginning of the whole llambda phages life system;
gateway to cloning technology:=-=============== Let's Start at the Beginning For lambda to integrate into bacterial chromosomes, as it does during lysogenization, two proteins catalyze the insertion of the phage DNA into the bacterial chromosome at a specific recombination site (att) present in the genome. The reverse reaction, excision of the phage genome from the E. coli chromosome, is mediated by three proteins—some viral, some bacterial. The presence or absence of a single protein, Xis, and the particular recombination sites involved, control the direction of these recombination reactions. These recombination proteins recognize four types of recombination sites. The B and P types sites are used for integration, and the L and R types for excision. GATEWAY Technology exploits modified versions of these att sites to do the cutting and pasting steps of the recombination reactions in vitro."....... www.bioresearchonline.com/article.mvc/GATEWAY-Cloning-TechnologyA-Universal-Cloning-0001#let=============== B P L R====== ....."The GATEWAY Technology recreates the lambda recombination reactions in vitro through a cocktail of recombination proteins and a set of vectors containing the att sequences they recognize. Two main pathways are involved. In one, the LR reaction, a cDNA or other DNA sequence flanked by attL sites is transferred by recombination from an Entry Clone into a Destination Vector, which contains attR sites. In this process, the Destination Vector conveys some functionally useful elements, such as a promoter, fusion tag, new replicon, or selection marker, to the final recombination product. The resulting molecule, called an Expression Clone, is a subclone of the starting DNA sequence, correctly positioned (same orientation and reading frame) in a new vector backbone. Life Technologies plans initially to offer nine types of Destination Vectors, for native protein and His6- or GST N-fusions in E. coli, insect cells, and mammalian cells. Additional GATEWAY-compatible vectors will continue to become available (for a myriad of applications). Ultimately, essentially any vector-containing technology can be compatible, allowing clones selected from one assay system to be "automatically" transferred into other systems for further analysis.:".................... ----- ....."The second GATEWAY pathway is the BP reaction, in essence the reverse of the LR reaction. The BP reaction transfers a DNA insert, flanked by 25 bp attB sites, from an Expression clone, into a vector donated by a plasmid containing attP sites. The final product is an Entry Clone containing the transferred DNA sequence. Using this approach, DNA segments can be transferred into and out of Entry Clones and Expression Clones at will, making the system completely open ended.
An important application of the BP reaction allows rapid, efficient, directional PCR Cloning. For this, a PCR product made using primers with 5'-terminal attB sites is recombined in the BP reaction with an attP plasmid to generate an Entry Clone of the PCR product. The resulting construct can be used to further transfer the cloned DNA into any number of Destination Vectors.".....Entry Clones also can be prepared with standard approaches using restriction enzyme and ligase. Life Technologies Inc. currently offers five different Entry Vectors for this purpose, providing an assortment of restriction sites for inserting your DNA sequence, or the presence or absence ribosome binding sites. Likewise, Entry Clone cDNA libraries can be prepared in an Entry Vector, something Life Technologies Inc. is likely to do in the future.4 ways to make Entry Clonesimages.vertmarkets.com/crlive/files/Images/816576D7-AE5C-11D3-9A76-00A0C9C83AFB/image3.jpg============== LR cloning reaction: images.vertmarkets.com/crlive/files/Images/816576D7-AE5C-11D3-9A76-00A0C9C83AFB/image4.jpg============== ......"The price for all this speed and convenience is that att sites flank your gene. Of the four kinds of att sites used in the GATEWAY Cloning Technology, only the attB sites that flank your gene in the Expression Clones are of any consequence. Fortunately, these are small (25 bp) and lack the hairpin structure of the loxP sites of the Cre/lox system (so expression levels remain high). The Life Technologies folks have taken out all the stop codons and made new specificities, so the attB sites are highly specific and don't have any noticeable effect on protein expression. In fact, translation across the attB sites doesn't even seem to change the in vivo protein-protein interactions of the yeast two-hybrid system.Beyond that, there are two recombination cocktails to keep track of, the BP CLONASE for cloning PCR products or moving your gene back out of an Expression Clone, and the LR CLONASE for moving your gene from an Entry Clone into any of the Destination Vectors. No cutting, no polishing, no phosphatasing, no gel purifications, no digests for orientation, etc., etc., etc. The reactions are so simple; they've been automated.
Any Legal Stuff to Worry About? Not for the academic researcher. Industrial types don't need to do anything up front (if doing research), and nothing at all until the volumes of CLONASE Enzyme Mix get pretty serious. Then there are licenses to arrange, but no royalties or reach through. There are licenses for commercial use of the system.www.bioresearchonline.com/article.mvc/GATEWAY-Cloning-TechnologyA-Universal-Cloning-0001#let Free range..........................Universal genes? View from the Laboratory To create a set of usable reagents representing an entire genome-worth of information, in this case, C. elegans, Vidal has turned to Life Technologies' Gateway technology. With this technology, he is creating a "macro-array" of all the open reading frames from his model organism—the ORFeome, they call it—which can then be produced in defined expression systems, the exact nature of which would vary depending on your purpose. You might choose yeast, for example, when you want to look at protein interactions through two-hybrid systems, or E. coli, when protein isolation is a goal.www.bioresearchonline.com/article.mvc/GATEWAY-Cloning-TechnologyA-Universal-Cloning-0001#letskyship
|
|
|
Post by skyship on Oct 1, 2010 17:08:37 GMT -5
ORFeome: new thread:
skyship
|
|