Difference between revisions of "Part:BBa K627006"

 
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<partinfo>BBa_K627006 short</partinfo>
 
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An appropriate vector containing the biobrick mdnA-myc-geneIII-fusion gene is a very important milestone of the development of a functional phage display system for screening of a microviridin (mdnA) library. Microviridins are tricyclic peptides from cyanocacteria which are able to bind and inhibit proteases. The gene III protein is a coat protein from the filamentous bacteriophage M13. It appears only 3-5 times on the tip of the phage and is responsible for infection of bacterial cells. After transformation of E. coli with the vector and co-infection with helper phages E. coli cells are able to produce phage particles carrying microviridin on their surface. Using these phages the fundamental suitability of phage display as a screening method for mdnA varieties was indicated. This has great importance for identifying microviridin varieties of therapeutical relevance. The inserted myc sequence enables the easy detection or purification . In our project the successful expression of the mdnA-myc -gene-III-fusion protein was determined by ELISA. Subsequent the production of phages carrying mdnA in E. coli was analyzed by phage display.
 
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K627006 SequenceAndFeatures</partinfo>
 
  
  
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The biobrick mdnA-myc-geneIII-fusion gene is important for testing the suitability of phage display system as an appropriate screening method for a recombinant mdnA-library.<br>
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===Functional Parameters===
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MdnA encoding the tricyclic peptide microviridin in cyanocacteria is known to be able to bind and inhibit proteases. Thus they have a high potential for therapy as they can block disease-relevant proteases. <br>
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<partinfo>BBa_K627006 parameters</partinfo>
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The gene III protein is a coat protein from the filamentous bacteriophage M13. It appears only 3-5 times on the tip of the phage and is responsible for infection of bacterial cells. In phage display genes from a DNA-library are usually fused to the gene III and displayed as fusion protein on the surface of the phages. Because it has been shown that proteins of interest fused to the carboxy-terminus of gene-III-proteins can be functionally displayed only the sequence for this part was used.<br>
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The inserted myc sequence enables the easy detection or purification.<br>
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After transformation of E. coli with a vector containing the mdnA-myc-geneIII-fusion gene and co-infection with helper phages E. coli cells were able to produce phage particles carrying mdnA on their surface. This was shown by ELISA test. Using these phages the fundamental suitability of phage display as a screening method for mdnA varieties was pointed.

Revision as of 23:06, 21 September 2011

Fusion part of mdnA gene (from mdn-cluster) with myc-tag and gene III


The biobrick mdnA-myc-geneIII-fusion gene is important for testing the suitability of phage display system as an appropriate screening method for a recombinant mdnA-library.
MdnA encoding the tricyclic peptide microviridin in cyanocacteria is known to be able to bind and inhibit proteases. Thus they have a high potential for therapy as they can block disease-relevant proteases.
The gene III protein is a coat protein from the filamentous bacteriophage M13. It appears only 3-5 times on the tip of the phage and is responsible for infection of bacterial cells. In phage display genes from a DNA-library are usually fused to the gene III and displayed as fusion protein on the surface of the phages. Because it has been shown that proteins of interest fused to the carboxy-terminus of gene-III-proteins can be functionally displayed only the sequence for this part was used.
The inserted myc sequence enables the easy detection or purification.
After transformation of E. coli with a vector containing the mdnA-myc-geneIII-fusion gene and co-infection with helper phages E. coli cells were able to produce phage particles carrying mdnA on their surface. This was shown by ELISA test. Using these phages the fundamental suitability of phage display as a screening method for mdnA varieties was pointed.