Expression of ECP-2 through quanitative real-time PCR
Poster Number
7
Format
Poster Presentation
Abstract/Artist Statement
Spiders produce a variety of silks that display extraordinary molecular and mechanical properties. These different silk types are composed of distinct proteins that are formed in seven specialized glands for typical orb-weaving spiders. Previously in the black widow spider Lactrodectus hesperus a novel gene encoding a protein that is part of egg case silk was isolated. Recent studies have shown that egg case silk protein (ECP-1) was expressed in the major ampullate (MA) gland as well as the tubuliform gland of the black widow spider. The gene encoding ECP-1 was isolated from a cDNA library using reverse genetic approaches. Current developments have shown the presence of another ‘sister’ gene called egg case protein-2 (ECP-2) which shows ~53% homology at the amino acid level to ECP-1. Using western blot analysis ECP-1 was shown to be expressed in the major ampullate and tubliform glands. Because we have yet to generate an antibody against ECP-2 we are using quantitative real-time PCR to measure the levels of mRNA in different silk glands. Although ECP-1 and ECP-2 have similar nucleotide sequences as well as amino acid sequences two different primer sets have been developed to differentiate ECP-2 from ECP-1 and will be used to examine ECP-2 expressed by quantitative real-time PCR. Since ECP-1 is produced in the MA and tubuliform glands ECP-2 is anticipated to be found in the same anatomical locations. Further research is being conducted in order to determine the glands responsible for ECP-2 production.
Location
Pacific Geosciences Center
Start Date
30-4-2005 1:00 PM
End Date
30-4-2005 3:00 PM
Expression of ECP-2 through quanitative real-time PCR
Pacific Geosciences Center
Spiders produce a variety of silks that display extraordinary molecular and mechanical properties. These different silk types are composed of distinct proteins that are formed in seven specialized glands for typical orb-weaving spiders. Previously in the black widow spider Lactrodectus hesperus a novel gene encoding a protein that is part of egg case silk was isolated. Recent studies have shown that egg case silk protein (ECP-1) was expressed in the major ampullate (MA) gland as well as the tubuliform gland of the black widow spider. The gene encoding ECP-1 was isolated from a cDNA library using reverse genetic approaches. Current developments have shown the presence of another ‘sister’ gene called egg case protein-2 (ECP-2) which shows ~53% homology at the amino acid level to ECP-1. Using western blot analysis ECP-1 was shown to be expressed in the major ampullate and tubliform glands. Because we have yet to generate an antibody against ECP-2 we are using quantitative real-time PCR to measure the levels of mRNA in different silk glands. Although ECP-1 and ECP-2 have similar nucleotide sequences as well as amino acid sequences two different primer sets have been developed to differentiate ECP-2 from ECP-1 and will be used to examine ECP-2 expressed by quantitative real-time PCR. Since ECP-1 is produced in the MA and tubuliform glands ECP-2 is anticipated to be found in the same anatomical locations. Further research is being conducted in order to determine the glands responsible for ECP-2 production.