Identification of Promoter Region of MaSp2 Protein

The goal of our project was to find the promoter region of the protein molecule MaSp2 from the black widow spider. We accomplished this task by performing polymerase chain reaction (PCR) with genomic DNA isolated from spider tissue and engineered oligonucleotides designed to amplify different fragments of DNA that contained the putative promoter region. Spider silks are known to have intriguing physical properties such as being lightweight, extremely strong and elastic. For a time, spider silk and its proteins, called spidroins, have been attempted to be massively produced as synthetic silks that mimic its versatility for use in commercial applications. But in order to do so, investigators must first unravel how spiders synthesize natural silk within their distinct silk glands and their precise mechanisms and processes. We were successful in amplifying the 5\u27 flanking DNA region of the MaSp2 gene (300 bp, 500 bp, 1000 bp, and 2000 bp regions). After amplification, we inserted these fragments into the pBAD-TOPO cloning vector and verified the presence of the putative promoter regions using restriction digestion analyses and agarose gel electrophoresis. These promoter fragments were also subject to DNA sequence analysis. Our long-term goal is to use these upstream regions to identify the endogenous promoter region ofMaSp2