DNA Isolation and Characterization of a novel fibroin from the black widow spider, Latrodectus hesperus
Poster Number
13
Format
Poster Presentation
Abstract/Artist Statement
Spider silk has been under intense research in recent years due its high tensile strength, extensibility and toughness. Silk also has several other desirable molecular features, including its biocompatibility and non-toxic nature. Scientists have taken particular interest in the expression, assembly, and spinning processes of silk to serve the environment in the future for a variety of different applications. Using a cDNA library prepared from the silk-producing glands of the black widow spider, Latrodectus hesperus, we searched the library for novel cDNAs that encoded for members of the spider silk family. Twenty-five different plaques were randomly isolated, amplified and their recombinant viral chromosomes were treated with a helper virus to release the plasmid carrying the cDNAs from the viral chromosome. After retrieval of the plasmids carrying the cDNAs from the library, we confirmed the presence of cDNAs in the cloning vectors by performing restriction digestion analysis. Following the validation of the presence of cDNA inserts, the plasmids were sent off for DNA sequencing. Using bioinformatics to analyze the DNA sequences of the cDNAs in the cloning vector, we found that one clone encoded a novel fibroin family member. This cloned contained the hallmark signatures for fibroins, including the conserved, non-repetitive C-terminus and internal repeats.
Location
DeRosa University Center, Ballroom B
Start Date
2-5-2009 1:00 PM
End Date
2-5-2009 3:00 PM
DNA Isolation and Characterization of a novel fibroin from the black widow spider, Latrodectus hesperus
DeRosa University Center, Ballroom B
Spider silk has been under intense research in recent years due its high tensile strength, extensibility and toughness. Silk also has several other desirable molecular features, including its biocompatibility and non-toxic nature. Scientists have taken particular interest in the expression, assembly, and spinning processes of silk to serve the environment in the future for a variety of different applications. Using a cDNA library prepared from the silk-producing glands of the black widow spider, Latrodectus hesperus, we searched the library for novel cDNAs that encoded for members of the spider silk family. Twenty-five different plaques were randomly isolated, amplified and their recombinant viral chromosomes were treated with a helper virus to release the plasmid carrying the cDNAs from the viral chromosome. After retrieval of the plasmids carrying the cDNAs from the library, we confirmed the presence of cDNAs in the cloning vectors by performing restriction digestion analysis. Following the validation of the presence of cDNA inserts, the plasmids were sent off for DNA sequencing. Using bioinformatics to analyze the DNA sequences of the cDNAs in the cloning vector, we found that one clone encoded a novel fibroin family member. This cloned contained the hallmark signatures for fibroins, including the conserved, non-repetitive C-terminus and internal repeats.