Identification of a New cDNA that Codes for Wrapping Silk in Black Widow Spiders
Poster Number
18
Format
Poster Presentation
Abstract/Artist Statement
Latrodectus hesperus produces silk with high tensile strength and extensibility. This allows it to be up to five times stronger than steel, when compared on a weight-to-strength basis. There are no products on the market based on black widow spider silk because of the complications associated with its synthetic production. There is much to be revealed about the natural production of black widow spider silk. The purpose of this study was to combine genetics and proteomics in order to learn more about the natural production of silk in L. hesperus. A cDNA library was constructed from the silk glands of black widow spiders. cDNAs were randomly selected from the library in hopes of discovering novel silk genes. Isolation of each cDNA was accomplished through single- clone excision to obtain recombinant plasmids, transformation of the vectors into E. coli, followed by double restriction digestion to verify the presence of cDNA inserts. Successfully isolated cDNAs were sequenced, and then translated into a protein sequence. Unique protein sequences were then searched against a list of peptides, obtained experimentally from the black widow spider silk using MS/MS analysis. Eighteen of the initial 25 cDNAs were successfully isolated and sequenced. One translated cDNA was found to contain a predicted peptide that matched a peptide obtained from MS/MS analysis. The discovery of the cDNA containing the peptide sequence is evidence of its existence in L. hesperus. Further study of this cDNA may reveal more information on its role in silk production.
Location
DeRosa University Center, Ballroom B
Start Date
2-5-2009 1:00 PM
End Date
2-5-2009 3:00 PM
Identification of a New cDNA that Codes for Wrapping Silk in Black Widow Spiders
DeRosa University Center, Ballroom B
Latrodectus hesperus produces silk with high tensile strength and extensibility. This allows it to be up to five times stronger than steel, when compared on a weight-to-strength basis. There are no products on the market based on black widow spider silk because of the complications associated with its synthetic production. There is much to be revealed about the natural production of black widow spider silk. The purpose of this study was to combine genetics and proteomics in order to learn more about the natural production of silk in L. hesperus. A cDNA library was constructed from the silk glands of black widow spiders. cDNAs were randomly selected from the library in hopes of discovering novel silk genes. Isolation of each cDNA was accomplished through single- clone excision to obtain recombinant plasmids, transformation of the vectors into E. coli, followed by double restriction digestion to verify the presence of cDNA inserts. Successfully isolated cDNAs were sequenced, and then translated into a protein sequence. Unique protein sequences were then searched against a list of peptides, obtained experimentally from the black widow spider silk using MS/MS analysis. Eighteen of the initial 25 cDNAs were successfully isolated and sequenced. One translated cDNA was found to contain a predicted peptide that matched a peptide obtained from MS/MS analysis. The discovery of the cDNA containing the peptide sequence is evidence of its existence in L. hesperus. Further study of this cDNA may reveal more information on its role in silk production.