title: [Expression and bioactive characterization of bacteriophage lysin gene of Bacillus anthracis in Escherichia coli].
abstract: The lysin gene of Bacillus anthracis-diagnosing bacteriophage, obtained by PCR amplification,was cloned into the Escherichia coli exepression vector pET22b which has been cleaved by EcoR I and Nde I. The recombinant vector pET22b-gamma lysin was verified to be correctly constructed by PCR, sequencing and enzyme digestion, and highly expressed in E. coli BL21 (DE3), which accounted for about 40 percent of total protein in E. coli BL21 (DE3), while in the 5L fermentor the expression level reached 15g/L. After expression, disruption and purification with three-step chromatography, Streamline SP, SP HP and Sephacryl S-100, the recombinant gamma lysin was finally obtained with purity of higher than 95 percent as determined by gel scan. The final yield following SP HP was 19.1 percent, with a greater-than-350-fold increase in specific activity. The pure enzyme has been shown active to Bacillus anthracis, and not to E. coli, Bacillus subtilis and Bacillus cereus. Its specific activity was about 1400 u/mg.
authors: Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors: Xiao-Jing
authors: XJ
authors: Li
authors: Hao
authors: H
authors: Zhang
authors: Xue-Qi
authors: XQ
authors: Fu
authors: Yan-Ying
authors: YY
authors: Li
authors: Jing
authors: J
authors: Chen
authors: Yu-Ling
authors: YL
authors: Li
authors: Hong-Qing
authors: HQ
authors: Fang
authors: Hui-Peng
authors: HP
authors: Chen
mesh_terms: D001408:Bacillus anthracis / Q000821:virology*; D001435:Bacteriophages / Q000201:enzymology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D014764:Viral Proteins