[Construction of the prokaryotic expression vector and expression of murine testis-specific gene Biot2].
OBJECTIVE
To construct a prokaryotic expression vector of mice gene Biot2, and to express the gene in E.coli/XL-Blue.


METHODS
The total RNA was extracted from mice testes tissues. Biot2 gene fragment was amplified by RT-PCR and was cloned into the pQE30 vector. The recombinant vector was identified by restriction endonuclease digestion analysis and DNA sequencing, and then it was transformed into E.coli/XL-Blue through IPTG induction to express the target protein bearing 6-His tag, which was analyzed by SDS-PAGE and Western blot.


RESULTS
After E.coli/XL-Blue was transformed with recombinant vector pQE30-Biot2 and induced with IPTG, the recombinant protein with relative molecular mass about 17.7 kD was obtained.


CONCLUSIONS
Recombinant expression vector pQE30-Biot2 is constructed and expressed successfully, which will be helpful to our further research.
Wang
Chun-ting
CT
State Key Laboratory of Biotherapy, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China. chtwang@163.com
Zhang
Peng
P
Peng
Feng
F
Yang
Han-shuo
HS
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism*
D000951:Antigens, Neoplasm; C543854:Ccdc7b protein, mouse; D011993:Recombinant Fusion Proteins