Klee H J, Gordon M P, Nester E W
J Bacteriol. 1982 Apr;150(1):327-31. doi: 10.1128/jb.150.1.327-331.1982.
A wide host range cosmid vector has been constructed by insertion of the lambda cos site into the plasmid pRK2501. This cosmid, which is maintained in Agrobacterium tumefaciens and is compatible with the Ti plasmid, has been used to make a clone bank of the A. tumefaciens pTiA6 plasmid. Several pTiA6 cosmids have been used to complement Tn5-induced Ti plasmid mutations. Five avirulent mutations which map outside of the region of the plasmid maintained in plant tumours (T-DNA) could be complemented in a trans orientation. Two mutations which are located on a single HpaI restriction fragment outside of the T-DNA, as well as three mutations which map within the T-DNA region, could not be complemented in a trans orientation in a REC- host.
通过将λ噬菌体粘性末端位点(cos位点)插入质粒pRK2501构建了一种广宿主范围的黏粒载体。这种黏粒可在根癌土壤杆菌中维持,并且与Ti质粒兼容,已被用于构建根癌土壤杆菌pTiA6质粒的克隆文库。几个pTiA6黏粒已被用于互补Tn5诱导的Ti质粒突变。位于植物肿瘤中维持的质粒区域(T-DNA)之外的五个无毒突变可以反式互补。位于T-DNA之外的单个HpaI限制片段上的两个突变以及位于T-DNA区域内的三个突变,在REC-宿主中不能反式互补。