Molecular Cloning of CAMK2A Gene

The role of calcium/calmodulin-dependent protein kinase II (CaMKIIα) as a signaling molecule inside neurons could influence function of the brain in learning and memory. Calcium signaling is crucial for several aspects of plasticity at glutamatergic synapses. CaMKIIα drives synaptic insertion of new receptor subunits and phosphorylates receptor subunits directly to enhance channel conductance. These processes are widely believed to play critical roles in the long-term regulation of synaptic transmission at the cellular level, and in learning and memory. The molecular cloning of the gene in mouse and rat could amplify and grow it. Sequencing results from the forward primer showed that the construct was present and it matched with the initial portion of the untranslated region, 3’UTR of CAMK2A gene of mouse. The reverse primer gave no sequence, probably because the Kanamycin portion got deleted from the construct. It was also seen that the end part of the sequence from forward primer had a portion of WPRE (Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element). This also validated the fact that Kanamycin portion got deleted. The sequencing results from the WPRE reverse primer had a major portion of the WPRE gene and then had a small portion of the CAMK2A 3’UTR. But this portion was from a middle portion of the UTR. This showed that the construct did not have the entire 3’UTR in it. The gel did not give any band anywhere after all three polymerase chain reactions, which showed that it did not get amplified. The translation regulation of the protein encoded by the gene could prove the long-term regulation of potentiation and depression and thus learning and memory.