Hunting For Novel Spider Silk Genes in the Black Widow Spider: Exploration of L. hesperus cDNA sequences
Poster Number
15
Format
Poster Presentation
Faculty Mentor Name
Craig Vierra
Abstract/Artist Statement
Spider silks have been a major focus in today's society due to their exemplary mechanical properties such as high tensile strength and flexibility. Much of the research on spider silks has focused on how to mass-produce silk fibers for industrial applications. Through a series of techniques and procedures, our research hopes to discover new silk genes that encode silk proteins. Using a eDNA library constructed from the silk-producing glands of the black widow spider, Latrodectus hesperus, we randomly selected over 125 different recombinant viruses carrying potentially distinct cDNAs. We performed single clone excisions from the different recombinant viruses and transformed the recovered phagemid particles into E. coli. Plasmids were retrieved and digested with restriction enzymes to analyze the size of the eDNA inserts using agarose gel electrophoresis. Plasmids carrying eDNA inserts from the screen were then sent to the University of Florida for DNA sequence analysis. We then compared our nucleic acid sequences against sequences deposited in the GenBank® database using the computer program BLAST. Results will be discussed. Our long-term goal is to identify new silk genes, as well as use the known eDNA sequences to print on the surface of glass slides using robotics to create "spider DNA chips". These spider DNA chips will then be used to monitor global changes in gene expression profiles during the silk process of spiders to understand more regarding the in vivo production of spider silk.
Location
Wendell Phillips Center, 1st floor hallways
Start Date
3-5-2008 1:00 PM
End Date
3-5-2008 3:00 PM
Hunting For Novel Spider Silk Genes in the Black Widow Spider: Exploration of L. hesperus cDNA sequences
Wendell Phillips Center, 1st floor hallways
Spider silks have been a major focus in today's society due to their exemplary mechanical properties such as high tensile strength and flexibility. Much of the research on spider silks has focused on how to mass-produce silk fibers for industrial applications. Through a series of techniques and procedures, our research hopes to discover new silk genes that encode silk proteins. Using a eDNA library constructed from the silk-producing glands of the black widow spider, Latrodectus hesperus, we randomly selected over 125 different recombinant viruses carrying potentially distinct cDNAs. We performed single clone excisions from the different recombinant viruses and transformed the recovered phagemid particles into E. coli. Plasmids were retrieved and digested with restriction enzymes to analyze the size of the eDNA inserts using agarose gel electrophoresis. Plasmids carrying eDNA inserts from the screen were then sent to the University of Florida for DNA sequence analysis. We then compared our nucleic acid sequences against sequences deposited in the GenBank® database using the computer program BLAST. Results will be discussed. Our long-term goal is to identify new silk genes, as well as use the known eDNA sequences to print on the surface of glass slides using robotics to create "spider DNA chips". These spider DNA chips will then be used to monitor global changes in gene expression profiles during the silk process of spiders to understand more regarding the in vivo production of spider silk.