绵羊肺炎支原体甘油代谢酶GlpO的表达、鉴定与纯化  

Expression,identification and purification of glycerol metabolism enzyme GlpO from Mycoplasma ovipneumoniae

作　　者：马臣杰[1,2] 侯玉婷[1,2] 李敏[1,2] 何玉龙 何萍[1,2] 张斯民[1,2] 张文清[1,2] 

机构地区：[1]西部特色生物资源保护与利用教育部重点实验室,银川750021 [2]宁夏大学生命科学学院,银川750021

出　　处：《黑龙江畜牧兽医》2014年第12期1-3,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目"绵羊肺炎支原体甘油代谢酶GLPO功能研究"(31260035)

摘　　要：为了研究绵羊肺炎支原体GlpO蛋白的结构和功能,试验构建了绵羊肺炎支原体Y98株甘油代谢酶GlpO的重组表达载体p ET-32a-glp O,并在大肠杆菌BL21(DE3)中对其进行表达,然后经SDS-PAGE和Western-blot对表达产物进行鉴定,最后利用Ni柱在AKTA系统上对GlpO进行纯化。结果表明:重组表达的GlpO蛋白在SDS-PAGE中的分子质量与GlpO理论分子质量54 ku接近,并且能够被anti-S-Tag antibody特异性识别。说明本试验成功在大肠杆菌中可溶性表达了绵羊肺炎支原体GlpO蛋白。To study the structure and function of glycerol metabolism enzyme （GlpO） from Mycoplasma ovipneumoniae, the recombinant expresion vector pET - 32a - glpO of glycerol metabolism enzyme GlpO from the Y98 strain of Mycoplasma ovipneumoniae was constructed. It was in- duced to express GlpO in Escherichia coli BL21 （DE3） by IPTG, and then the expression product was identified by SDS -PAGE and Western blotting. The GlpO was purified on the AKTA system using the Ni column. The results showed that the molecular mass of the recombinant GlpO protein was close to its theoretical molecular mass of 54 kD by SDS - PAGE, and it could be recognized by the specificity of the anti - S - Tag antibody. The results indicate that the GlpO protein Of Mycoplasma ovipneumoniae was successfully expressed in soluble form in Escherichia coli.

关 键 词：绵羊肺炎支原体 甘油代谢酶 重组表达 大肠杆菌 蛋白纯化 

分 类 号：S858.26[农业科学—临床兽医学] S852.62[农业科学—兽医学]