摘要
目的构建巴西诺卡菌P61基因原核重组表达载体,在大肠杆菌中表达具有生物活性的P61蛋白,为P61蛋白的进一步相关研究提供实验基础。方法通过基因合成的方法合成P61基因,将其插入到质粒pET30a(+)载体并导入BL21大肠杆菌,应用IPTG进行诱导表达。通过Western Blot对表达产物进行分析。应用过氧化氢酶检测试剂盒检测其过氧化氢酶活性。结果P61蛋白在大肠杆菌中可溶性表达,且具有较高的过氧化氢酶活性,能被感染巴西诺卡菌的小鼠血清识别。结论成功构建了巴西诺卡菌P61蛋白的原核表达质粒,并能在原核表达宿主E.coli BL21中进行高效表达,且表达产物具有过氧化氢酶活性。
We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and in-duced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N,brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.
作者
吉兴照
唐璐
侯雪新
孙丽娜
韦超
徐帅
司晨琛
李振军
JI Xing-zhao;TANG Lu;HOU Xue-xin;SUN Li-na;WEI Chao;XU Shuai;SI Chen-chen;LI Zhen-jun(Institute for Communicable Disease Prevention and Control,Stat-Key Laboratory for Communicable Disease Prevention and Control Chinese Center for Disease Control and prevention,Beijing 102206,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2017年第3期260-263,共4页
Chinese Journal of Zoonoses
基金
863课题(No.2014AA021404)
国家科技重大专项(No.2013ZXl0004221)
公益性卫生行业专项(No.201302006)联合资助
