Construction of pain-related fusion gene plasmids and inhibitory effect of small interfering RNA

Objective: To study the inhibitory effects of small hairpin RNA（shRNA） expression vector on expression of the Cav2.2e37a gene. Methods: Expression plasmid EGFP-e37a/e37b was constructed.BHK cell line was co-transfected with the expression plasmid EGFP-e37a and level of EGFP-e37a fusion protein was quantified by fluorescent microscopy and fluorescence activated cell sorting（FACS）. Results: The clones were identified by PCR and DNA sequence analysis.The results were shown as expected.siRNAII could inhibit 90% of EGFP-e37a expression. Conclusion: The EGFP-e37a/e37b fusion gene plasmids can be constructed and expressed in BHK cell line,and RNA interference（RNAi） can effectively reduce the levels of e37a transcripts and protein expression,thus providing a fresh approach to screening the efficient small interfering RNA（siRNA）.