宿主介导的结核分枝杆菌中PvuII限制修饰
Host-mediated modification of PvuII restriction in Mycobacterium tuberculosis.
作者信息
van Soolingen D, de Haas P E, Blumenthal R M, Kremer K, Sluijter M, Pijnenburg J E, Schouls L M, Thole J E, Dessens-Kroon M W, van Embden J D, Hermans P W
机构信息
Laboratory for Bacteriology and Antimicrobial Agents, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
出版信息
J Bacteriol. 1996 Jan;178(1):78-84. doi: 10.1128/jb.178.1.78-84.1996.
Restriction endonuclease PvuII plays a central role in restriction fragment length polymorphism analysis of Mycobacterium tuberculosis complex isolates with IS6110 as a genetic marker. We have investigated the basis for an apparent dichotomy in PvuII restriction fragment pattersn observed among strains of the M. tuberculosis complex. The chromosomal regions of two modified PvuII restriction sites, located upstream of the katG gene and downstream of an IS1081 insertion sequence, were studied in more detail. An identical 10-bp DNA sequence (CAGCTGGAGC) containing a PvuII site was found in both regions, and site-directed mutagenesis analysis revealed that this sequence was a target for modification. Strain-specific modification of PvuII sites was identified in DNA from over 80% of the nearly 800 isolates examined. Furthermore, the proportion of modifying and nonmodifying strains differs significantly from country to country.
限制性内切酶PvuII在以IS6110作为遗传标记的结核分枝杆菌复合群分离株的限制性片段长度多态性分析中起着核心作用。我们研究了在结核分枝杆菌复合群菌株中观察到的PvuII限制性片段模式明显二分法的基础。对位于katG基因上游和IS1081插入序列下游的两个修饰的PvuII限制性位点的染色体区域进行了更详细的研究。在两个区域中都发现了一个相同的包含PvuII位点的10碱基DNA序列（CAGCTGGAGC），并且定点诱变分析表明该序列是修饰的靶点。在近800株被检测菌株中，超过80%的菌株DNA中鉴定出PvuII位点的菌株特异性修饰。此外，修饰菌株和未修饰菌株的比例在不同国家之间存在显著差异。
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