[Construction of the prokaryotic expression vector and expression of murine testis-specific gene Biot2]

Abstract

Aim: 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.

Conclusion: Recombinant expression vector pQE30-Biot2 is constructed and expressed successfully, which will be helpful to our further research.