关键词: 结核分枝杆菌； 异柠檬酸裂解酶； 基因表达

Abstract:

Isocitrate lyase(ICL) gene was amplified by polymerase chain reaction (PCR) from template Mycobacterium tuberculosis H₃₇R
v strain genomic DNA and was cloned into expression vector pET28b, named pET28b ICL. The recombined plasmid pET28bICL was transformed into E.coli BL21(DE3). The optimum conditions for ICL expression in E.coli BL21 (DE3) were determined to be 025 mmol/L IPTG induction for 4 h at 20 ℃. The expressed ICL was further purified by means of NiNTA resin affinity chromatography. The recombinant ICL was purified in a highly active state with a specific activity of 24 μmol/(mg·min).

Key words: Mycobacterium tuberculosis, isocitrate lyase, gene expression