Isolation of cDNAs from Lactrodectus hesperus
Poster Number
31
Format
Poster Presentation
Abstract/Artist Statement
For the past years, experiments to obtain genes that code for proteins spun into spider silk fibers have been carefully conducted. The silk produced by Latrodectus hesperus, commonly known as the black widow spider, is known for its high tensile strength and durability, and thus by manufacturing it as synthetic biological product, it would be possible to replace standard engineering materials such as steel. In our studies, we attempted to identify new silk genes that coded for proteins spun into spider silk fibers. Moreover, we also searched our library for gene products involved in the silk pathway. For our experimental work, we were given a cDNA library that was prepared from the abdominal silk-producing glands of the L. hesperus. To isolate different cDNA molecules from the library, we plated the library and randomly picked 25 individual plaques. Spider cDNAs were excised from the viral chromosome and the resulting recombinant plasmids were then transformed into the Escherichia coli. The plasmids were then isolated from the bacteria. In order to confirm that cDNAs were present within the recovered plasmids, the plasmid was digested with restriction enzymes and their products were subject to agarose gel electrophoresis. After confirmation the plasmids were subject to DNA sequencing and bioinformatics was used to study the identity of the retrieved genes.
Location
DeRosa University Center, Ballroom B
Start Date
1-5-2010 1:00 PM
End Date
1-5-2010 3:00 PM
Isolation of cDNAs from Lactrodectus hesperus
DeRosa University Center, Ballroom B
For the past years, experiments to obtain genes that code for proteins spun into spider silk fibers have been carefully conducted. The silk produced by Latrodectus hesperus, commonly known as the black widow spider, is known for its high tensile strength and durability, and thus by manufacturing it as synthetic biological product, it would be possible to replace standard engineering materials such as steel. In our studies, we attempted to identify new silk genes that coded for proteins spun into spider silk fibers. Moreover, we also searched our library for gene products involved in the silk pathway. For our experimental work, we were given a cDNA library that was prepared from the abdominal silk-producing glands of the L. hesperus. To isolate different cDNA molecules from the library, we plated the library and randomly picked 25 individual plaques. Spider cDNAs were excised from the viral chromosome and the resulting recombinant plasmids were then transformed into the Escherichia coli. The plasmids were then isolated from the bacteria. In order to confirm that cDNAs were present within the recovered plasmids, the plasmid was digested with restriction enzymes and their products were subject to agarose gel electrophoresis. After confirmation the plasmids were subject to DNA sequencing and bioinformatics was used to study the identity of the retrieved genes.