Philipp W J, Nair S, Guglielmi G, Lagranderie M, Gicquel B, Cole S T
Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, Paris, France.
Microbiology (Reading). 1996 Nov;142 ( Pt 11):3135-45. doi: 10.1099/13500872-142-11-3135.
A Dral restriction map of the approximately 4.35 Mb circular chromosome of the vaccine strain Mycobacterium bovis BCG Pasteur was constructed by linking all 21 Dral fragments, ranging in size from 6 to 820 kb, using specific clones that spanned the Dral recognition sites as hybridization probes. The positions of 20 known genes were also established. Comparison of the resultant genome map with that of the virulent tubercle bacillus Mycobacterium tuberculosis H37Rv revealed extensive global conservation of the genomes of these two members of the M. tuberculosis complex. Possible sites of evolutionary rearrangements were localized on the chromosome of M. bovis BCG Pasteur by comparing the Asnl restriction profile with that of M. tuberculosis H37Rv. When selected cosmids from the corresponding areas of the genome of M. tuberculosis H37Rv were used as hybridization probes to examine different BCG strains, wild-type M. bovis and M. tuberculosis H37Rv, a number of deletions up to 10 kb in size, insertions and other polymorphisms were detected. In addition to the known deletions covering the genes for the protein antigens ESAT-6 and mpt64, other genetic loci exhibiting polymorphisms or rearrangements were detected in M. bovis BCG Pasteur.
通过使用跨越DraI识别位点的特定克隆作为杂交探针,将大小从6至820 kb不等的所有21个DraI片段连接起来,构建了牛分枝杆菌卡介苗巴斯德疫苗株约4.35 Mb环状染色体的DraI限制酶图谱。还确定了20个已知基因的位置。将所得基因组图谱与强毒结核杆菌结核分枝杆菌H37Rv的图谱进行比较,发现结核分枝杆菌复合群这两个成员的基因组在整体上具有广泛的保守性。通过比较牛分枝杆菌卡介苗巴斯德株与结核分枝杆菌H37Rv的AsnI限制酶切图谱,确定了进化重排的可能位点。当使用来自结核分枝杆菌H37Rv基因组相应区域的选定黏粒作为杂交探针,检测不同的卡介苗菌株、野生型牛分枝杆菌和结核分枝杆菌H37Rv时,发现了一些大小达10 kb的缺失、插入及其他多态性。除了已知的覆盖蛋白抗原ESAT-6和mpt64基因的缺失外,在牛分枝杆菌卡介苗巴斯德株中还检测到了其他表现出多态性或重排的基因位点。