Analysis and Search of Silk Gene Proteins in Latrodectus Hesperus
Poster Number
17
Format
Poster Presentation
Abstract/Artist Statement
Latrodectus hesperus is commonly known as the black widow spider. The different silk fibers it produces are extensively studied to enable large-scale commercial applications. Its soluble nature, high tensile strength, toughness, and biodegradability provide promising advancements and improvements to current products. However, many crucial aspects of the silks and their properties have yet to be discovered or fully analyzed to allow these applications. In hopes of obtaining new silk genes that could be utilized to produce synthetic silk in bacteria or yeast, a cDNA library was created from the silk-producing glands of the black widow spider. The cDNA library was then plated out and random recombinant viruses were isolated. The amplification, excision, and digestion of plasmid cloning vector embedded in the viral chromosome was then carried out, isolating the individual plasmids carrying the cDNAs for DNA sequencing. With the use of translational programs, protein BLAST algorithms, nucleotide BLAST programs, the protein sequences and their identities could be determined. Translated cDNA sequences were then compared to a list of generated peptide sequences obtained from mass spectrometry. Identical matches showed that the translated cDNAs were actuality responsible for making that specific protein found in the fiber. Through this process, the cDNAs encoding Masp1,Masp2, and an unidentified wrap protien were identified.
Location
DeRosa University Center, Ballroom B
Start Date
2-5-2009 1:00 PM
End Date
2-5-2009 3:00 PM
Analysis and Search of Silk Gene Proteins in Latrodectus Hesperus
DeRosa University Center, Ballroom B
Latrodectus hesperus is commonly known as the black widow spider. The different silk fibers it produces are extensively studied to enable large-scale commercial applications. Its soluble nature, high tensile strength, toughness, and biodegradability provide promising advancements and improvements to current products. However, many crucial aspects of the silks and their properties have yet to be discovered or fully analyzed to allow these applications. In hopes of obtaining new silk genes that could be utilized to produce synthetic silk in bacteria or yeast, a cDNA library was created from the silk-producing glands of the black widow spider. The cDNA library was then plated out and random recombinant viruses were isolated. The amplification, excision, and digestion of plasmid cloning vector embedded in the viral chromosome was then carried out, isolating the individual plasmids carrying the cDNAs for DNA sequencing. With the use of translational programs, protein BLAST algorithms, nucleotide BLAST programs, the protein sequences and their identities could be determined. Translated cDNA sequences were then compared to a list of generated peptide sequences obtained from mass spectrometry. Identical matches showed that the translated cDNAs were actuality responsible for making that specific protein found in the fiber. Through this process, the cDNAs encoding Masp1,Masp2, and an unidentified wrap protien were identified.