重组结核分枝杆菌Mr38000蛋白的表达纯化和鉴定医学.docVIP

重组结核分枝杆菌Mr38000蛋白的表达纯化和鉴定医学 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：重组结核分枝杆菌Mr38000蛋白的表达纯化和鉴定医学 1 2：诱导菌超声裂解后沉淀; 3: 诱导菌超声裂解后上清; 6 3 讨论 6 文2：结核分枝杆菌RpfA蛋白的表达纯化与鉴定 7 1材料和方法 8 参考文摘引言： 12 原创性声明（模板） 14 文章致谢（模板） 14 正文 重组结核分枝杆菌Mr38000蛋白的表达纯化和鉴定医学 文1：重组结核分枝杆菌Mr38000蛋白的表达纯化和鉴定医学 Cloning, Expression and Purification of Mr 38 000 protein from Mycobacterium tuberculosis ZHANG Ming， LI Jin-cheng， LIN Hang (Department of internal neurology, 2. emergency department, the Fu Zhou general hospital, Fu Zhou, 350025, P. R. China) [Abstract] To obtain Mycobacterium tuberculosis Mr 38 000 protein from H37Rv-DNA, express efficiently in E. coli and purify the Mr 38 000 protei. Mr 38 000 protein gene was amplified by PCR from the genome of H37Rv and cloned into pGEM-T-Easy vector. After sequenced, Mr 38 000 protein gene was recombinated expression vector pQE-80L by restriction enzyme digestion, named pQE-80L-Mr 38 000. The plasmid pQE-80L-Mr 38 000 was traformed into DH5α and induced by IPTG. Mr 38 000 protein expression was analyzed by SDSand confirmed by western blot with mice-specific mAb agait (His)6. Mr 38 000 protein gene was identical with GeBank reported. The pQE-80L-Mr 38 000 vector expressed protein with relative molecule weight about kD, which could be caught by (His)6 mAb. The expressed protein could be purified by Ni-NTA system kit with denative condition. Mr 38 000 protein gene had been successfully cloned and efficiently expressed in , The results established the basis for further study of the function of Mr 38 000 protein. [Key words] Mr38 000 protein; expression; purification; Mycobacterium tuberculosis (MTB) 结核分枝杆菌（Mycobacterium tuberculosis, MTB）是结核病(tuberculosis, TB)的病原体。其在患者体内主要为细胞内生长。诊断方法的欠缺是结核病流行的重要原因。 目前常用的PPD试验，常使BCG疫苗接种者被误检为阳性[1]，且对后期结核患者敏感性较低，存在明显不足。近年来， 有研究发现Mr 38 000 蛋白具有强的免疫原性，而成为结核分枝杆菌抗原的研究热点[2, 3]，可能成为结核病血清学诊断试剂和疫苗研究的候选抗原之一。为此，我们在原核表达载体中表达结核分枝杆菌 Mr 38 000蛋白并对其进行了纯化，为进一步研究其抗原性和主要功能提供方便。 １　材料和方法 材料 　MTB毒株H37Rv、 DH5α由本室保存; pGEM-T-Easy及Wi