Expression and purification of Rv0073 gene of Mycobacterium tuberculosis in Escherichia coli

• DOI: 10.3760/cma.j.issn.1673-4394.2015.03.006
• Publication Date: 2015-05-05
• Authors: Gaoxiang Sun; Xinling Pan; Jiayi Cui; Hong Ling
• Keywords: Inclusion bodies

Objective To construct a prokaryotic expression vector carrying Mycobacterium tuberculosis(MTB)Rv0073 gene and express the recombinant protein in Escherichia coli(E.coli).

Methods The Rv0073 from MTB H37Rv strain was amplified by polymerase chain reaction (PCR) cloned into plasmid pET26b(+ ). Then, E.coli BL21 transformed with plasmid. The induced using Isopropyl β-D-1-thiogalactopyranoside (IPTG)at 25 ℃, 28 30 34 ℃ 37 respectively. IPTG induction for 2 h, 4 h 7 also examined. Finally, we purified MagneHis™ Ni-Parties kit.

Results We constructed plasmid, pET26b-Rv0073. successfully detected SDS-PAGE.Induction temperature did not affect expression. But time is related to of decrease increasing temperature.The formed inclusion bodies after We could purify protein.

Conclusion The will become basis further production studying on biological functions protein.

Key words: Mycobacterium tuberculosis; Rv0073; Expression; Purification