Title

The search for the gene sequence of dragline silk from the Major Ampullate Spidroin II gland in the black widow spider (Latrodectus hesperus)

Poster Number

31

Format

Poster Presentation

Abstract/Artist Statement

Each type of silk from the black widow spider (Lactrodectus Hesperus) have different functions. We expect that the gene differences of each type of silk will lead to differences in their material properties. The goal of our research is to find the molecular gene sequence for the dragline silk of the Major Ampullate Spidroin II gland (MaSp2) in the black widow spider. A cDNA library was constructed using the DNA of MaSp2 glands because of their high composition of silk mRNA's* 10%. Primers were designed to accommodate the degeneracy of the DNA code at a silk region that is evolutionarily thought to be a common motif amongst other spider species. Three PCR reactions were run with forward and reverse primers, forward primers only, and reverse primers only with an annealing temperature ranging from 45-52 degrees Celsius for each. Using gel electrophoresis, three distinct bands were found for the PCR product with forward and reverse primers. The three gene pieces were each inserted into a cloning vector and transformed into E. colt. Plasmid vectors were isolated and sequenced. A positive clone was found with high homology to previously reported silk genes and its relevance will be discussed.

Location

Pacific Geosciences Center

Start Date

20-4-2002 9:00 AM

End Date

20-4-2002 5:00 PM

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Apr 20th, 9:00 AM Apr 20th, 5:00 PM

The search for the gene sequence of dragline silk from the Major Ampullate Spidroin II gland in the black widow spider (Latrodectus hesperus)

Pacific Geosciences Center

Each type of silk from the black widow spider (Lactrodectus Hesperus) have different functions. We expect that the gene differences of each type of silk will lead to differences in their material properties. The goal of our research is to find the molecular gene sequence for the dragline silk of the Major Ampullate Spidroin II gland (MaSp2) in the black widow spider. A cDNA library was constructed using the DNA of MaSp2 glands because of their high composition of silk mRNA's* 10%. Primers were designed to accommodate the degeneracy of the DNA code at a silk region that is evolutionarily thought to be a common motif amongst other spider species. Three PCR reactions were run with forward and reverse primers, forward primers only, and reverse primers only with an annealing temperature ranging from 45-52 degrees Celsius for each. Using gel electrophoresis, three distinct bands were found for the PCR product with forward and reverse primers. The three gene pieces were each inserted into a cloning vector and transformed into E. colt. Plasmid vectors were isolated and sequenced. A positive clone was found with high homology to previously reported silk genes and its relevance will be discussed.