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B:2型多杀性巴氏杆菌分离株特异独特序列的克隆

Cloning of a unique sequence specific to isolates of type B:2 Pasteurella multocida.

作者信息

Townsend K M, Dawkins H J, Zeng B J, Watson M W, Papadimitriou J M

机构信息

Department of Pathology, University of Western Australia.

出版信息

Res Vet Sci. 1996 Nov;61(3):199-205. doi: 10.1016/s0034-5288(96)90063-6.

Abstract

Two Carter type B Pasteurella multocida isolates, Izatnagar 52 and 25, isolated from cases of haemorrhagic septicaemia (HS), were used in a modified subtractive hybridisation technique with the specific aim of cloning unique DNA sequences related to the pathogenesis of HS. Biochemical and protein analyses have shown these isolates to be similar, but reports indicate that they have differences in pathogenicity. The subtracted inserts were screened against genomic DNA from a wide range of P multocida isolates, with two distinct fragments demonstrating specific hybridisation with Carter type B isolates that cause HS. No identity was observed with either Carter type E isolates or non-HS type B strains. The clones were sequenced and a search of the GenBank database revealed significant identity of the clone A3b (296 nt) to P haemolytica lipoprotein, whereas there was no significant identity with 6b (956 nt). Both these fragments had a high level of identity (72.8 to 76.9 per cent) to the H influenzae Rd genome.

摘要

从出血性败血症(HS)病例中分离出的两株卡特B型多杀性巴氏杆菌菌株,即伊扎特纳加尔52和25,被用于一种改良的消减杂交技术,其具体目的是克隆与HS发病机制相关的独特DNA序列。生化和蛋白质分析表明这些分离株相似,但报告显示它们在致病性上存在差异。用来自多种多杀性巴氏杆菌分离株的基因组DNA对消减后的插入片段进行筛选,有两个不同的片段与引起HS的卡特B型分离株呈现特异性杂交。未观察到与卡特E型分离株或非HS型B菌株有同源性。对这些克隆进行测序,并在GenBank数据库中搜索,发现克隆A3b(296 nt)与溶血巴氏杆菌脂蛋白有显著同源性,而与6b(956 nt)没有显著同源性。这两个片段与流感嗜血杆菌Rd基因组有高度同源性(72.8%至76.9%)。

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