Nakata N, Matsuoka M, Kashiwabara Y, Okada N, Sasakawa C
Leprosy Research Center, National Institute of Health, Higashimurayama-shi, Tokyo, Japan.
J Bacteriol. 1997 May;179(9):3053-7. doi: 10.1128/jb.179.9.3053-3057.1997.
Synthetic oligonucleotide primers based on the DNA sequence data of the Escherichia coli, Mycobacterium tuberculosis, and Mycobacterium intracellulare katG genes encoding the heme-containing enzyme catalase-peroxidase were used to amplify and analyze the Mycobacterium leprae katG region by PCR. A 1.6-kb DNA fragment, which hybridized to an M. tuberculosis katG probe, was obtained from an M. leprae DNA template. Southern hybridization analysis with a probe derived from the PCR-amplified fragment showed that the M. leprae chromosome contains only one copy of the putative katG sequence in a 3.4-kb EcoRI-BamHI DNA segment. Although the nucleotide sequence of the katG region of M. leprae was approximately 70% identical to that of the M. tuberculosis katG gene, no open reading frame encoding a catalase-peroxidase was detectable in the whole sequence. Moreover, two DNA deletions of approximately 100 and 110 bp were found in the M. leprae katG region, and they seemed to be present in all seven M. leprae isolates tested. These results strongly suggest that M. leprae lacks a functional katG gene and catalase-peroxidase activity.
基于编码含血红素的过氧化氢酶-过氧化物酶的大肠杆菌、结核分枝杆菌和胞内分枝杆菌katG基因的DNA序列数据,合成寡核苷酸引物用于通过聚合酶链反应(PCR)扩增和分析麻风分枝杆菌katG区域。从麻风分枝杆菌DNA模板中获得了一个与结核分枝杆菌katG探针杂交的1.6kb DNA片段。用PCR扩增片段衍生的探针进行Southern杂交分析表明,麻风分枝杆菌染色体在一个3.4kb的EcoRI - BamHI DNA片段中仅包含一份推定的katG序列拷贝。尽管麻风分枝杆菌katG区域的核苷酸序列与结核分枝杆菌katG基因的序列约70%相同,但在整个序列中未检测到编码过氧化氢酶-过氧化物酶的开放阅读框。此外,在麻风分枝杆菌katG区域发现了两个约100和110bp的DNA缺失,并且它们似乎存在于所有测试的7株麻风分枝杆菌分离株中。这些结果强烈表明麻风分枝杆菌缺乏功能性的katG基因和过氧化氢酶-过氧化物酶活性。