Katarina Conradson, Shane Turgeon, Nicole Benson, Ian Walbridge, Paula Croonquist
The objective of this experiment was to clone and sequence the GAPDH encoding gene GAPC for sequencing in purple prairie clover. The central question was to investigate whether the GAPC gene is conserved in the Dalea purpurea, as in other plants. It was hypothesized that the GAPC gene is found in Dalea purpurea, and that although it constitutes a unique sequence not previously reported, it shares a high degree of homology with other cytosolic GAPDH encoding genes in other plant species. In this experiment, plant cells were lysed, and DNA was extracted, purified, and run through PCR to amplify the GAPC gene. E. coli was used to clone the gene of interest, which was then extracted. Gel electrophoresis was utilized twice, after PCR reactions and digesting extracted vectors, to separate and visualize amplified gene product. The PCR reactions did not yield any PCR products, and previously amplified PCR products were ligated into the vector. The cloned vectors were extracted from the transformed bacteria and digested to separate the gene insert from the vector. The electrophoresis gel of the vector digest did contain bands consistent with a successful gene insert. It was concluded that samples contained the GAPC gene insert, thus indicating successful isolation and cloning of this gene in purple prairie clover.