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 Cav2.2e[37a] 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).