Mullany P, Clayton C L, Pallen M J, Slone R, al-Saleh A, Tabaqchali S
Department of Medical Microbiology, St. Bartholomew's Hospital Medical College, West Smithfield, London, UK.
FEMS Microbiol Lett. 1994 Nov 15;124(1):61-7. doi: 10.1111/j.1574-6968.1994.tb07262.x.
Screening of a Clostridium difficile lambda EMBL3 gene library with antisera raised against C. difficile culture supernatant identified several clones expressing a 31-kDa protein. A 1.8-kb HindIII fragment subcloned from one of the clones was sufficient for expression of the 31-kDa polypeptide. Southern blot analysis showed a region homologous to this fragment to be present in all of 13 different C. difficile strains tested. Sequence analysis of the 1.8-kb fragment revealed three adjacent open reading frames. A database search showed that these three open reading frames appeared to encode homologues of three consecutive enzymes in the butanol/butyrate-producing pathway of Clostridium acetobutylicum (crotonase, beta-hydroxybutyryl coenzyme A dehydrogenase and thiolase).
用针对艰难梭菌培养上清液产生的抗血清筛选艰难梭菌λEMBL3基因文库，鉴定出几个表达31 kDa蛋白的克隆。从其中一个克隆亚克隆的1.8 kb HindIII片段足以表达31 kDa多肽。Southern印迹分析表明，在测试的所有13种不同艰难梭菌菌株中均存在与该片段同源的区域。对1.8 kb片段的序列分析揭示了三个相邻的开放阅读框。数据库搜索表明，这三个开放阅读框似乎编码丙酮丁醇梭菌丁醇/丁酸产生途径中三种连续酶的同源物（巴豆酸酶、β-羟基丁酰辅酶A脱氢酶和硫解酶）。
