Post by aqt on Feb 8, 2010 17:38:57 GMT -5
What are fusion proteins and why are they important?
If a protein of interest is present in very small quantities insufficient to characterize, to raise an antibody against, or to utilize for therapeutic purposes, recombinant techniques are utilized. By fusing a known protein or peptide DNA sequence with the DNA of the protein of interest, fusion proteins (a combination of the protein of interest tagged with the known protein or peptide) can be produced in culture by microorganisms in large quantities.
How are these Pro-Fusion™ products used?
When researchers are attempting to produce a protein of interest, a DNA sequence which codes for the protein of interest is tagged or fused with the sequence for another protein (e.g. MBP or GFP) or a sequence that codes for an identifiable peptide (e.g. HA or c-Myc). This recombinant DNA is then introduced into a microorganism, which expresses the protein of interest as well as the tagged protein or peptide.
To isolate or to localize the protein of interest, the tag, which is now part of that protein, is isolated or localized, since that tag is attached to the protein of interest. The Fusion-Aid™ kits are used to isolate these "fusion proteins". The fusion protein binds to the appropriate agarose-bound antibody spin column and is thus separated from all of the other proteins in the culture supernatant or cell lysate. The biotinylated anti-MBP, anti-GFP, anti-HA, and anti-c-Myc are used to follow the production of the fusion proteins or to identify the fusion protein in a western blot format. If the fusion protein is expressed in tissues, the biotinylated antibodies can be used to localize, within the tissue section, the protein of interest.
Protein Tagging Technology:
GFP and MBP are two proteins commonly used to produce fusion proteins. The DNA sequences for these proteins are attached to the DNA sequence of the protein of interest. When the organism expresses the protein of interest, the protein "tags" (e.g. GFP or MBP) are also produced. Since these tags are now a part of the expressed protein, anti-GFP or anti-MBP can be employed to localize or isolate the protein of interest.
Epitope Tagging Technology:
In a manner similar to that described above for GFP or MBP, a DNA sequence coding for a peptide is introduced into the DNA sequence of the protein of interest. When expressed, this fusion protein contains the epitope of the tag defined by the peptide amino acid sequence. The agarose bound anti-HA or anti-c-Myc is used to isolate the fusion protein from the culture supernatant or the cell lysate. The biotinylated antibodies are used to identify the proteins of interest in western blots or potentially to localize the fusion proteins in the tissue.
www.vectorlabs.com/canada/products.asp?catID=390
If a protein of interest is present in very small quantities insufficient to characterize, to raise an antibody against, or to utilize for therapeutic purposes, recombinant techniques are utilized. By fusing a known protein or peptide DNA sequence with the DNA of the protein of interest, fusion proteins (a combination of the protein of interest tagged with the known protein or peptide) can be produced in culture by microorganisms in large quantities.
How are these Pro-Fusion™ products used?
When researchers are attempting to produce a protein of interest, a DNA sequence which codes for the protein of interest is tagged or fused with the sequence for another protein (e.g. MBP or GFP) or a sequence that codes for an identifiable peptide (e.g. HA or c-Myc). This recombinant DNA is then introduced into a microorganism, which expresses the protein of interest as well as the tagged protein or peptide.
To isolate or to localize the protein of interest, the tag, which is now part of that protein, is isolated or localized, since that tag is attached to the protein of interest. The Fusion-Aid™ kits are used to isolate these "fusion proteins". The fusion protein binds to the appropriate agarose-bound antibody spin column and is thus separated from all of the other proteins in the culture supernatant or cell lysate. The biotinylated anti-MBP, anti-GFP, anti-HA, and anti-c-Myc are used to follow the production of the fusion proteins or to identify the fusion protein in a western blot format. If the fusion protein is expressed in tissues, the biotinylated antibodies can be used to localize, within the tissue section, the protein of interest.
Protein Tagging Technology:
GFP and MBP are two proteins commonly used to produce fusion proteins. The DNA sequences for these proteins are attached to the DNA sequence of the protein of interest. When the organism expresses the protein of interest, the protein "tags" (e.g. GFP or MBP) are also produced. Since these tags are now a part of the expressed protein, anti-GFP or anti-MBP can be employed to localize or isolate the protein of interest.
Epitope Tagging Technology:
In a manner similar to that described above for GFP or MBP, a DNA sequence coding for a peptide is introduced into the DNA sequence of the protein of interest. When expressed, this fusion protein contains the epitope of the tag defined by the peptide amino acid sequence. The agarose bound anti-HA or anti-c-Myc is used to isolate the fusion protein from the culture supernatant or the cell lysate. The biotinylated antibodies are used to identify the proteins of interest in western blots or potentially to localize the fusion proteins in the tissue.
www.vectorlabs.com/canada/products.asp?catID=390