1.Prokryotic expression of CFP-10 protein of Mycobacterium tuberculosis
Shisheng FENG ; Jianqin LIANG ; Jinhe WANG ; Xueqiong WU
Journal of Chinese Physician 2013;(3):324-327
Objective To construct recombinant plasmid containing CFP-10 gene of Mycobacterium tuberculosis(MTB).Methods The gene fragment of CFP-10 was amplified by PCR from Mycobacterium tuberculosis H37Rv genomic DNA and cloned to pET-32a(+) vector.The recombinant plasmid pET-32a-CFP-10 was transformed into E.coli BL21 (DE3) and induced by IPTG.Results CFP-10 gene fragment was amplified from genomic DNA of Mycobacterium tuberculosis H37Rv strain,and thepET-32a(+) prokaryotic recombinant plasmid was constructed successfully.The recombinant protein was expressed with the induction of IPTG.Conclusions The prokaryotic expression vector for CFP-10 was successfully constructed and the recombinant protein was highly expressed in E.coli BL21 (DE3),which lays a foundation for its subsequent immunological function study.
2.Effects of genistein on the proliferation of cardiac fibroblasts
Zhan GAO ; Miaozhang ZHU ; Shisheng ZHOU ; Shunyan LU ; Haitao GUO ; Feng GAO ; Xinliang MA
Chinese Journal of Pharmacology and Toxicology 2001;15(2):159-160
To study the effect of genistein on the proliferation of cardiac fibroblasts(CF), CFs were cultured from neonatal rat hearts, DNA synthesis of the cells was determined by incorporation of [3H]TdR into DNA, the cell cycle was measured by flow cytometric analysis. Genistein(0.5-50 μmol*L-1) attenuated 2.5% fetal calf serum-induced proliferation of CF in concentration-dependent manner. Genistein(50 μmol*L-1) arrested CF cell progression at G2/M phase. The results suggest that genistein be a potential substance for treatment of cardiac fibrosis.