Whittington R, Marsh I, Choy E, Cousins D
NSW Agriculture, Elizabeth Macarthur Agricultural Institute, Menangle, New South Wales, Australia.
Mol Cell Probes. 1998 Dec;12(6):349-58. doi: 10.1006/mcpr.1998.0194.
IS1311 is an insertion sequence from Mycobacterium avium and M. avium subsp. paratuberculosis. Using a 180 bp fragment of IS 1311 as a probe, 7-10 copies of IS1311 were revealed in strains of M. avium subsp. paratuberculosis. With a given restriction enzyme, the restriction fragment length polymorphism patterns obtained from isolates of M. avium subsp. paratuberculosis from cattle were all identical, but they differed from those of isolates from sheep, which could be separated into two types. A 1259 bp fragment of IS1311 produced by polymerase chain reaction (PCR) from two isolates of M. avium subsp.paratuberculosis from cattle and two isolates from sheep was sequenced and compared to the sequence known from M. avium. Apart from five point differences the sequences were identical. Restriction endonuclease analysis (REA) of the PCR product was used to distinguish isolates of M. avium subsp. paratuberculosis from M. avium, in addition to the conventional test for IS900. In isolates of M. avium subsp. paratuberculosis from cattle the IS1311 gene was polymorphic at position 223, which enabled isolates from sheep and cattle to be distinguished by PCR-REA. These simple tests will be used to enhance disease control programmes for Johne's disease in ruminants. The findings of this study raise interesting questions about the evolution of M. avium subsp. paratuberculosis.
IS1311是来自鸟分枝杆菌和副结核分枝杆菌亚种的一种插入序列。以IS1311的180 bp片段为探针，在副结核分枝杆菌亚种菌株中发现了7 - 10个IS1311拷贝。用一种特定的限制性内切酶，从牛的副结核分枝杆菌分离株获得的限制性片段长度多态性模式完全相同，但与绵羊分离株的模式不同，绵羊分离株可分为两种类型。对来自牛的两株副结核分枝杆菌分离株和来自绵羊的两株分离株通过聚合酶链反应（PCR）产生的1259 bp的IS1311片段进行测序，并与鸟分枝杆菌已知序列进行比较。除了五个位点差异外，序列相同。除了传统的IS900检测外，PCR产物的限制性内切酶分析（REA）用于区分副结核分枝杆菌亚种和鸟分枝杆菌的分离株。在来自牛的副结核分枝杆菌亚种分离株中，IS1311基因在223位是多态性的，这使得可以通过PCR - REA区分来自绵羊和牛的分离株。这些简单的检测将用于加强反刍动物约翰氏病的疾病控制计划。本研究的结果提出了关于副结核分枝杆菌亚种进化的有趣问题。
