Isolation of novel glue proteins from black widow egg case silk
Poster Number
12
Format
Poster Presentation
Abstract/Artist Statement
The peptides obtained from a tryptic digestion of egg case silk were analyzed, and it was observed that the masses of the fragments corresponded with the masses of peptides that were posttranslationally modified with GlcNAc or GalNAc residues, which indicated the presence of glycoproteins in the silk glands from which they were taken. The partial peptide sequence...SDGGSNVGGNEYR was used to design primers for the purpose of amplifying the gene which encodes the peptides of these glycoproteins. After running PCR using one anchor primer and one gene-specific primer, several DNA bands of various sizes were discovered. These were taken as candidate cDNA fragments and they were extracted and purified from the gel, then ligated into a Topo vector for bacterial transformation. Colonies grown on LB-AMP plates were then cultured and plasmid DNA was isolated. The DNA was then sent for sequencing, and their sequences were analyzed using bioinformatics. The results will be discussed.
Location
Pacific Geosciences Center
Start Date
5-5-2007 1:00 PM
End Date
5-5-2007 3:00 PM
Isolation of novel glue proteins from black widow egg case silk
Pacific Geosciences Center
The peptides obtained from a tryptic digestion of egg case silk were analyzed, and it was observed that the masses of the fragments corresponded with the masses of peptides that were posttranslationally modified with GlcNAc or GalNAc residues, which indicated the presence of glycoproteins in the silk glands from which they were taken. The partial peptide sequence...SDGGSNVGGNEYR was used to design primers for the purpose of amplifying the gene which encodes the peptides of these glycoproteins. After running PCR using one anchor primer and one gene-specific primer, several DNA bands of various sizes were discovered. These were taken as candidate cDNA fragments and they were extracted and purified from the gel, then ligated into a Topo vector for bacterial transformation. Colonies grown on LB-AMP plates were then cultured and plasmid DNA was isolated. The DNA was then sent for sequencing, and their sequences were analyzed using bioinformatics. The results will be discussed.