中文摘要：

从结核分枝杆菌H37Rv基因组中扩增出ESAT-6基因,克隆入pET21a（＋）载体。将成功构建的pET21a（＋）-ESAT-6重组质粒转化入感受态BL21（DE3）中,经诱导表达后,使用SDS-PAGE和Western blot鉴定。超声破碎后发现目的蛋白以可溶性表达形式存在,经过Ni-NTA柱和DEAE-SepharoseTMFast Flow柱纯化,获得纯度约为95%的重组ESAT-6蛋白。将目的蛋白和RAW264.7细胞共孵育后,使用免疫荧光技术发现ESAT-6蛋白能够直接和RAW264.7的细胞膜结合。

英文摘要：

ESAT-6 gene was amplified by PCR from the genome of Mycobacterium tuberculosis H37Rv,and then cloned into pET21a（＋） plasmid.The recombinant plasmid that was successfully constructed was transformed into E.coli BL21（DE3）.After induced with IPTG,the expressed recombinant protein was confirmed by SDS-PAGE and Western blot.The vector yielded satisfactory levels of recombinant ESAT-6 protein expressed as a soluble protein in E.coli.After ultrasonication,the recombinant ESAT-6 protein was firstly purified by a column packed with Ni-NTA Resin and then a column packed with DEAE-SepharoseTM Fast Flow matrix.The purity of the purified protein was about 95%.The purified ESAT-6 protein was incubated with RAW264.7 cells,and the result got by Immunofluorescence showed that ESAT-6 could directly bind to the macrophage membrane.