Unwebbing the Unforeseen: Searching for new spider silk genes
Poster Number
33
Format
Poster Presentation
Abstract/Artist Statement
Spider silk is spun from a liquid protein solution. Silk fibers are about 1 micrometer in diameter. Despite spider silks thinness, it possesses high strength and elasticity. Due to its remarkable physical properties, spider silk has the potential to be used as an artificial material for bulletproof vests, parachute cords, surgical sutures and replacement ligaments. So far, seven distinct spider silk gene family members have been identified. In search for new silk genes, black widow spider cDNAs were isolated, amplified and sequenced. The library was initially constructed by inserting spider cDNAs into bacteriophage chromosomes to create a special cloning vector that is commonly referred to as a “pop-out” vector system. In order to retrieve individual spider cDNAs for DNA sequencing, we excised plasmids, along with their cDNAs, out of the recombinant viral chromosome using a single clone excision methodology. Following excision, cDNA-containing plasmids were transformed into XLOLR E. coli cells. Individual transformants were selected and grown overnight, followed by isolation of plasmid DNA using a standard miniprep DNA isolation procedure. In order to verify the presence of cDNA inserts, plasmids were digested with restriction enzymes to release cDNA inserts, followed by separation of the DNA fragments using agarose gel electrophoresis. Plasmids with cDNA inserts were sent for DNA sequence analysis. After completion of the DNA sequencing, we will analyze the sequences using computational biology to determine whether we have identified genes involved in the silk process.
Location
DeRosa University Center, Ballroom B
Start Date
1-5-2010 1:00 PM
End Date
1-5-2010 3:00 PM
Unwebbing the Unforeseen: Searching for new spider silk genes
DeRosa University Center, Ballroom B
Spider silk is spun from a liquid protein solution. Silk fibers are about 1 micrometer in diameter. Despite spider silks thinness, it possesses high strength and elasticity. Due to its remarkable physical properties, spider silk has the potential to be used as an artificial material for bulletproof vests, parachute cords, surgical sutures and replacement ligaments. So far, seven distinct spider silk gene family members have been identified. In search for new silk genes, black widow spider cDNAs were isolated, amplified and sequenced. The library was initially constructed by inserting spider cDNAs into bacteriophage chromosomes to create a special cloning vector that is commonly referred to as a “pop-out” vector system. In order to retrieve individual spider cDNAs for DNA sequencing, we excised plasmids, along with their cDNAs, out of the recombinant viral chromosome using a single clone excision methodology. Following excision, cDNA-containing plasmids were transformed into XLOLR E. coli cells. Individual transformants were selected and grown overnight, followed by isolation of plasmid DNA using a standard miniprep DNA isolation procedure. In order to verify the presence of cDNA inserts, plasmids were digested with restriction enzymes to release cDNA inserts, followed by separation of the DNA fragments using agarose gel electrophoresis. Plasmids with cDNA inserts were sent for DNA sequence analysis. After completion of the DNA sequencing, we will analyze the sequences using computational biology to determine whether we have identified genes involved in the silk process.