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目的克隆结核分支杆菌ｒｐｏＢ基因。方法以结核分支杆菌ｒｐｏＢ基因保守区的聚合酶链反应（ＰＣＲ）扩增片段为探针从已建立的结核分支杆菌基因文库中筛选ｒｐｏＢ基因。结果引物ＴＲ１、ＴＲ２ｂ扩增Ｈ３７ＲａＤＮＡ获１约４１１ｂｐ片段。经克隆及序列分析表明该片段与结核分支杆菌ｒｐｏＢ基因中央区域的序列同源。对约１２０００个克隆进行筛选，共获７个可疑阳性克隆，经过再次杂交证实其中３个克隆为阳性。其中１个克隆携带约３．８ｋｂ插入片段。一系列内切酶酶切分析后得出该３．８ｋｂ克隆片段的部分限制性内切酶酶谱。进一步分析表明所用４１１ｂｐ探针同源序列距两端分别为１ｋｂ和２．８ｋｂ。结论与报道的结核分支杆菌ｒｐｏＢ基因开放阅读框架比较，我们克隆的３．８ｋｂ片段包含结核分支杆菌ｒｐｏＢ完整基因的大部分，为进一步研究利福平及其它利福霉素类药物抗结核作用方式及耐药机制奠定了基础
Objective To clone Mycobacterium tuberculosis rpoB gene. Methods The rpoB gene was screened from the established Mycobacterium tuberculosis gene library by polymerase chain reaction (PCR) amplified fragment of the conserved region of Mycobacterium tuberculosis rpoB gene. Results Primers TR1, TR2b amplified H37RaDNA obtained a about 411bp fragment. Clone and sequence analysis showed that the fragment was homologous to the central region of Mycobacterium tuberculosis rpoB gene. About 12000 clones were screened, a total of 7 suspicious positive clones were obtained, of which 3 clones were confirmed positive by re-hybridization. One of the clones carries an approximately 3.8 kb insert. A series of endonuclease digestion analysis of the 3.8kb cloned fragment of the partial restriction enzyme zymogram. Further analysis indicated that the 411bp probe homology sequence used was 1 kb and 2.8 kb respectively from both ends. Conclusion Compared with the reported open reading frame of Mycobacterium tuberculosis rpoB gene, the cloned 3.8kb fragment contains most of the complete gene of Mycobacterium tuberculosis rpoB. To further study the anti-TB effect of rifampicin and other rifamycins And drug resistance mechanism laid the foundation