Construction of recombinant expression vector and prokaryotic expression of human epididymal sperm protein P34H / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To acquire purified recombinant human epididymal sperm protein P34H for basic and clinical studies.</p><p><b>METHODS</b>On the basis of cloning of P34H coding region, P34H fragment was subcloned into the pQE-30 expression vector. The recombinant expression vector designated pQE-30/P34H was transformed into E. coli to induce the expression of the recombinant protein P34H on the reduction of IPTG. After sonication, the recombinant protein P34H was purified from the supernatant with Ni-NTA resin under native conditions. It was identified by SDS-PAGE analysis and DNA sequencing.</p><p><b>RESULTS</b>Recombinant expression vector pQE-30/P34H was correctly constructed, identified with PCR and double-enzyme digestion. And the results of SDS-PAGE analysis and DNA sequencing showed that the protein was what we had hoped to acquire.</p><p><b>CONCLUSION</b>Purified recombinant P34H can be acquired successfully with the above mentioned prokaryotic expression method.</p>