Boivin R, Bellemare G, Dion P
Department of Phytology, Faculty of Agricultural and Food Sciences, Laval University, Québec, Canada.
Curr Microbiol. 1994 Jan;28(1):41-7. doi: 10.1007/BF01575984.
Narrow-host-range vectors, based on an indigenous replicon and containing a multiple cloning site, have been constructed in a Pseudomonas host capable of growth on unusual substrates. The new cloning vectors yield sufficient amounts of DNA for preparative purposes and belong to an incompatibility group different from that of the incP and incQ broad-host-range vectors. One of these vectors, named pDB47F, was used to clone, directly in Pseudomonas, DNA fragments from Agrobacterium, Pseudomonas, and Rhizobium. A clone containing Agrobacterium and KmR gene sequences was transformed with a higher efficiency than an RSF1010-derived vector (by as much as 1250-fold) in four out of five Pseudomonas strains tested. The considerable efficiency obtained with this system makes possible the direct cloning and phenotypic selection of foreign DNA in Pseudomonas.
基于本地复制子并含有多克隆位点的窄宿主范围载体，已在能够在特殊底物上生长的假单胞菌宿主中构建。这些新的克隆载体可产生足够量的DNA用于制备目的，并且属于与IncP和IncQ广宿主范围载体不同的不相容群。其中一个载体，命名为pDB47F，用于直接在假单胞菌中克隆来自农杆菌、假单胞菌和根瘤菌的DNA片段。在所测试的五株假单胞菌菌株中，有四株含有农杆菌和KmR基因序列的克隆，其转化效率比RSF1010衍生载体高（高达1250倍）。该系统获得的显著效率使得在假单胞菌中直接克隆和表型选择外源DNA成为可能。
