Bartosik D, Białkowska A, Baj J, Włodarczyk M
Department of Bacterial Genetics, Warsaw University, Poland.
Acta Microbiol Pol. 1997;46(4):387-92.
Two mobilizable cloning vectors, designated pABW1 and pAWB2, were constructed basing on the E. coli vector pBGS18 and oriT originating from RK2. In pABW2 the kanamycin resistance gene was replaced by a novel tetracycline resistance cassette derived from Tn1721. Both vectors, specific for E. coli, allow to perform the cloning steps in E. coli and then to efficiently transfer the constructs by conjugation to the host of choice. A vector which cannot propagate in the given host can be applied for identification of the host specific plasmid replicator regions. With the use of pABW2 we defined the minimal replicator region of pTAV202-a mini-derivative of the large pTAV1 plasmid of P. versutus. We also proved that RepC' encoded on this fragment is the principal initiator replication protein and that oriV is located along its coding sequence.
构建了两个可移动的克隆载体，分别命名为pABW1和pAWB2，它们基于大肠杆菌载体pBGS18和源自RK2的oriT构建而成。在pABW2中，卡那霉素抗性基因被一个源自Tn1721的新型四环素抗性盒所取代。这两个载体均对大肠杆菌具有特异性，能够在大肠杆菌中进行克隆步骤，然后通过接合作用将构建体高效转移至选定的宿主中。一个不能在给定宿主中繁殖的载体可用于鉴定宿主特异性质粒复制子区域。利用pABW2，我们确定了pTAV202（韦氏丙酸杆菌大质粒pTAV1的一个微型衍生物）的最小复制子区域。我们还证明了该片段上编码的RepC'是主要的起始复制蛋白，且oriV位于其编码序列上。
