非蛋白水解性B、E和F型肉毒梭菌的大亚基核糖体RNA基因及基因间隔区研究

Studies on the large subunit ribosomal RNA genes and intergenic spacer regions of non-proteolytic Clostridium botulinum types B, E and F.

作者信息

Campbell K D, East A K, Thompson D E, Collins M D

机构信息

Department of Microbiology, AFRC Institute of Food Research, Reading Laboratory, UK.

出版信息

Res Microbiol. 1993 Mar-Apr;144(3):171-80. doi: 10.1016/0923-2508(93)90042-z.

DOI:10.1016/0923-2508(93)90042-z

PMID:8210674

Abstract

The large-subunit ribosomal ribonucleic acid (23S rRNA) genes of non-proteolytic (group II) strains of Clostridium botulinum toxin types B, E and F were amplified using the polymerase chain reaction (PCR), and cloned in Escherichia coli. Sequence determination showed that the 23S rRNA genes were 2910 nucleotides in length, and comparative analysis revealed approximately 99.5% sequence similarity. The 23S rRNA gene sequence of a strain phenotypically resembling non-proteolytic C. botulinum, except in not producing botulinal neurotoxin, was also determined and displayed 99.5% sequence similarity with those from toxigenic strains. A diagnostic sequence within the 23S rRNA characteristic for non-proteolytic C. botulinum was identified and used for the design of an oligonucleotide probe. Molecular hybridizations with PCR-amplified rDNA targets provided a precise and reliable method of identifying non-proteolytic (or Group II) C. botulinum and closely related non-toxigenic strains.

摘要

利用聚合酶链反应（PCR）扩增了肉毒梭菌B、E和F型非蛋白水解性（II组）菌株的大亚基核糖体核糖核酸（23S rRNA）基因，并将其克隆到大肠杆菌中。序列测定表明，23S rRNA基因长度为2910个核苷酸，比较分析显示序列相似性约为99.5%。还测定了一株表型类似于非蛋白水解性肉毒梭菌，但不产生肉毒神经毒素的菌株的23S rRNA基因序列，其与产毒菌株的序列相似性为99.5%。在非蛋白水解性肉毒梭菌的23S rRNA中鉴定出一个诊断序列，并用于设计寡核苷酸探针。与PCR扩增的rDNA靶标的分子杂交提供了一种精确可靠的方法，用于鉴定非蛋白水解性（或II组）肉毒梭菌及密切相关的非产毒菌株。