[Construction of recombinant expression vector and prokaryotic expression of human epididymal sperm protein P34H].
OBJECTIVE
To acquire purified recombinant human epididymal sperm protein P34H for basic and clinical studies.


METHODS
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.


RESULTS
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.


CONCLUSIONS
Purified recombinant P34H can be acquired successfully with the above mentioned prokaryotic expression method.
Xia
Xinyi
X
Laboratory of Reproduction & Genetics, Nanjing General Hospital of Nanjing Command, PLA, Nanjing, Jiangsu 210002, China.
Huang
Yufeng
Y
Gao
Yun
Y
Zhu
Zhaoping
Z
D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D011994:Recombinant Proteins / Q000096:biosynthesis; D013401:Sugar Alcohol Dehydrogenases / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D013401:Sugar Alcohol Dehydrogenases; C023300:L-xylulose reductase