Title

Fishing for spider silk genes

Poster Number

51

Format

Poster Presentation

Abstract/Artist Statement

Spider silk is a remarkable material with respect to its tensile strength, ductility, and toughness. While remaining biodegradable, it is able to stretch to up to 140% its length and can even hold its strength down to forty degrees Celsius! We hypothesize that random selection of viruses from a cDNA library constructed in viral chromosomes will lead to the identification of new cDNA sequences that code for silk proteins. In order to test our hypothesis, we first isolated and amplified single recombinant viruses carrying different spider cDNAs. We used the method of single clone excision in vivo to excise plasmids containing the spider cDNAs from the viral chromosomes. The excised products, which were referred to as phagemid particles, were then transformed into a special strain of E. coli known as XLOLR. After selecting individual bacterial colonies carrying the plasmids and growing these cells to saturation, plasmid DNA was isolated and subject to restriction digestion. Digested products were analyzed using agarose gel electrophoresis to verify the presence of cDNA inserts. Following validation of inserts, the cDNAs will be analyzed by DNA sequencing and their corresponding sequences subject to bioinformatic analyses to determine the identity of the cDNAs and relevance to the silk pathway.

Location

DeRosa University Center, Ballroom B

Start Date

1-5-2010 1:00 PM

End Date

1-5-2010 3:00 PM

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May 1st, 1:00 PM May 1st, 3:00 PM

Fishing for spider silk genes

DeRosa University Center, Ballroom B

Spider silk is a remarkable material with respect to its tensile strength, ductility, and toughness. While remaining biodegradable, it is able to stretch to up to 140% its length and can even hold its strength down to forty degrees Celsius! We hypothesize that random selection of viruses from a cDNA library constructed in viral chromosomes will lead to the identification of new cDNA sequences that code for silk proteins. In order to test our hypothesis, we first isolated and amplified single recombinant viruses carrying different spider cDNAs. We used the method of single clone excision in vivo to excise plasmids containing the spider cDNAs from the viral chromosomes. The excised products, which were referred to as phagemid particles, were then transformed into a special strain of E. coli known as XLOLR. After selecting individual bacterial colonies carrying the plasmids and growing these cells to saturation, plasmid DNA was isolated and subject to restriction digestion. Digested products were analyzed using agarose gel electrophoresis to verify the presence of cDNA inserts. Following validation of inserts, the cDNAs will be analyzed by DNA sequencing and their corresponding sequences subject to bioinformatic analyses to determine the identity of the cDNAs and relevance to the silk pathway.