Isolation of cDNAs from Black Widow cDNA Library

Spider silk is extraordinarily flexible, elastic, and lightweight, but it is also extremely strong. Because spider silk is also biodegradable and can be produced without pollution, if it could be synthetically made, some possible uses are body armor, parachutes, and ropes, but the possibilities are virtually endless. Because of its unique properties, we isolated cDNA from a Black Widow cDNA library to find novel silk genes or other components involved in silk production. By plating plasmids containing the cDNA library in a viral chromosome, we were able to separate individual viruses, each containing a distinct cDNA sequence. Using single clone in vivo excision, plasmids inside the phage were excised and then transformed into E.coli. Transformants were then selected and grown in liquid cultures. Using plasmid miniprep purification, the cDNA was isolated. Restriction double digestion with restriction enzymes EcoRI and XhoI allowed for release of the cDNA from the cloning vectors, and analysis was performed using gel electrophoresis and DNA sequencing. Results will be discussed after the DNA sequencing reactions are analyzed using bioinformatic approaches