2nd Annual Academic Symposium - Katelyn St. Clair

Two important topics in cellular and molecular biology are the transfer of genetic information and biotechnology. In a series of laboratory exercises, students learn the importance of both by using a recombinant DNA clone to illustrate replication via PCR, transcription/translation, PAGE, and Western Blotting techniques. The previously used recombinant had several problems, including cross reactivity with pre-existing proteins in the translation reaction. The purpose of this project was to clone the Calpain 2 gene (CAPN2) to replace the previous recombinant and develop methods to avoid cross reactivity. A mouse CAPN2 cDNA was directionally cloned from skeletal muscle into an expression vector. The expected insert size of 2200bp was verified by restriction enzyme digestion and PCR. Protein of the expected size (78kDa) was synthesized from the CAPN2 recombinant using Transcend tRNA, which allows detection of newly synthesized protein with a colorimetric assay. These improved methods will replace those previously used.