When the Going Gets Tough, the Silk Keeps Going
Poster Number
32
Format
Poster Presentation
Abstract/Artist Statement
Spider silk has extraordinary material properties. The goal of this project was to isolate novel cDNAs from the black widow spider that code for proteins that are part of silk fibers or the silk assembly pathway. These novel genes may prove instrumental in producing new kinds of silk, or provide insight into ways in which silk may be synthetically mass-produced for military applications, where strength is much needed for combat equipment. In order to search for new silk genes, we plated a cDNA library prepared from the silk-producing glands of the spider to help separate the different silk genes from one another. Twenty-five individual viruses from the library were plugged, amplified and subject to single clone in vivo excision using the ExAssist helper phage. After excision, the phagemid particles, which contained virally-excised plasmids carrying the cDNAs, were transformed into E. coli. Then, in order to achieve adequate amounts of plasmid DNA for downstream studies, colonies from the transformants were selected and grown to saturation in liquid cultures. Saturated cultures were used to isolate plasmid DNA for restriction digestion analysis, followed by inspection of the DNA fragments using agarose gel electrophoresis. Plasmids determined to contain cDNA inserts were then subject to DNA sequence analysis. The results will be discussed after the cDNAs are sequenced and examined using bioinformatics software such as BLASTn and the ExPASy protein translation tool.
Location
DeRosa University Center, Ballroom B
Start Date
1-5-2010 1:00 PM
End Date
1-5-2010 3:00 PM
When the Going Gets Tough, the Silk Keeps Going
DeRosa University Center, Ballroom B
Spider silk has extraordinary material properties. The goal of this project was to isolate novel cDNAs from the black widow spider that code for proteins that are part of silk fibers or the silk assembly pathway. These novel genes may prove instrumental in producing new kinds of silk, or provide insight into ways in which silk may be synthetically mass-produced for military applications, where strength is much needed for combat equipment. In order to search for new silk genes, we plated a cDNA library prepared from the silk-producing glands of the spider to help separate the different silk genes from one another. Twenty-five individual viruses from the library were plugged, amplified and subject to single clone in vivo excision using the ExAssist helper phage. After excision, the phagemid particles, which contained virally-excised plasmids carrying the cDNAs, were transformed into E. coli. Then, in order to achieve adequate amounts of plasmid DNA for downstream studies, colonies from the transformants were selected and grown to saturation in liquid cultures. Saturated cultures were used to isolate plasmid DNA for restriction digestion analysis, followed by inspection of the DNA fragments using agarose gel electrophoresis. Plasmids determined to contain cDNA inserts were then subject to DNA sequence analysis. The results will be discussed after the cDNAs are sequenced and examined using bioinformatics software such as BLASTn and the ExPASy protein translation tool.