Identification of Novel cDNAs that Code for Spider Silk in Black Widow Spiders
Poster Number
39
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 learned about the natural production of black widow spider silk. The purpose of this study was to combine genetics and proteomics in order to understand 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 predicted protein sequences. These protein sequences were then searched against a database of black widow spider peptides, obtained experimentally by sequencing peptides using MS/MS analysis obtained from solubilized fibers digested with trypsin; these fibers were collected from the spider’s web, egg sac and attachment disc threads. Approximately 24 cDNAs were successfully isolated and subject to DNA sequence analysis. Further study of these cDNAs may reveal more information on their role in silk production.
Location
DeRosa University Center, Ballroom B
Start Date
1-5-2010 1:00 PM
End Date
1-5-2010 3:00 PM
Identification of Novel cDNAs that Code for Spider 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 learned about the natural production of black widow spider silk. The purpose of this study was to combine genetics and proteomics in order to understand 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 predicted protein sequences. These protein sequences were then searched against a database of black widow spider peptides, obtained experimentally by sequencing peptides using MS/MS analysis obtained from solubilized fibers digested with trypsin; these fibers were collected from the spider’s web, egg sac and attachment disc threads. Approximately 24 cDNAs were successfully isolated and subject to DNA sequence analysis. Further study of these cDNAs may reveal more information on their role in silk production.