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基于复制子pRRI7的杂交质粒从大肠杆菌向拟杆菌属和普雷沃菌属菌株的转移。

Transfer of hybrid plasmids based on the replicon pRRI7 from Escherichia coli to Bacteroides and Prevotella strains.

作者信息

Béchet M, Pheulpin P, Flint H J, Martin J, Dubourguier H C

机构信息

Laboratoire de Microbiologie, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.

出版信息

J Appl Bacteriol. 1993 May;74(5):542-8.

PMID:8486560
Abstract

New shuttle vectors based on a Prevotella ruminicola 9.5 kb cryptic plasmid (pRRI7) inserted within the Escherichia coli vector pKC71, carrying the Ccr/Emr Bacteroides marker, were constructed. These constructs (pKBR23-1 and pKBR23-2) were transferred into Bacteriodes distasonis, Bacteroides thetaiotaomicron, Bacteroides uniformis and into P. ruminicola NCFB 2202 either by conjugal mobilization or by electroporation. Another pRRI7 derivative based on pKC72, pKBR23-3, was smaller (13.1 kb) and non-mobilizable. By electroporation, it was transferred to Bact. distasonis and P. ruminicola. Being derived from pRRI7 which is compatible with the shuttle plasmid pRRI207, the host/vector combination involving P. ruminicola NCFB 2202 and pKBR23-3 offers new possibilities for genetic investigations in rumen anaerobic bacteria after further introduction of a second readily selectable marker within pRRI207 or pKBR23-3.

摘要

构建了基于插入大肠杆菌载体pKC71内的普氏栖瘤胃菌9.5 kb隐蔽质粒(pRRI7)的新型穿梭载体,该载体携带Ccr/Emr拟杆菌标记。这些构建体(pKBR23 - 1和pKBR23 - 2)通过接合转移或电穿孔法转入解脲拟杆菌、多形拟杆菌、单形拟杆菌以及普氏栖瘤胃菌NCFB 2202。另一个基于pKC72的pRRI7衍生物pKBR23 - 3较小(13.1 kb)且不可移动。通过电穿孔法,它被转入解脲拟杆菌和普氏栖瘤胃菌。由于源自与穿梭质粒pRRI207兼容的pRRI7,在pRRI207或pKBR23 - 3中进一步引入第二个易于选择的标记后,涉及普氏栖瘤胃菌NCFB 2202和pKBR23 - 3的宿主/载体组合为瘤胃厌氧菌中的基因研究提供了新的可能性。

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