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首页> 外文期刊>Journal of bacteriology >Gene transfer system for Rhodopseudomonas viridis.
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Gene transfer system for Rhodopseudomonas viridis.

机译：绿假单胞菌的基因转移系统。

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A gene transfer system for Rhodopseudomonas viridis was established which uses conjugation with Escherichia coli S17-I as the donor and mobilizable plasmids as vectors. Initially, plasmids of the incompatibility group P1 (pRK290 and pRK404) were used. The more effective shuttle vectors between E. coli and R. viridis, pKV1 and pKVS1, were derived from plasmid pBR322 and showed the highest conjugation frequency (10(-2] thus far demonstrated in purple bacteria. It was also demonstrated that Rhizobium meliloti can be used as a donor for conjugation with R. viridis. From a genomic cosmid library of R. viridis constructed in the vector pHC79, clones that coded for subunits H (puh operon), L, M and cytochrome c (puf operon) of the photosynthetic reaction center were isolated and characterized. For linkage of the two operons on the genome, cosmids that overlapped with the operon-carrying clones were identified. The relative positions of the two operons could not be determined, but the operons must be more than 100 kilobase pairs apart. Thus, the genomic organization of the reaction center in R. viridis is different from that of Rhodobacter capsulatus, for which a distance of about 39 kilobase pairs was determined. From a spontaneous mutant of R. viridis that is resistant to the herbicide terbutryn, the puf operon was cloned in pKVS1 and transferred by conjugation into R. viridis wild-type cells. The resulting exconjugants were resistant to the herbicide, which demonstrated that the puf operon on pKVS1 constructions was functionally expressed in R. viridis.
机译：建立了绿假单胞菌的基因转移系统，该系统使用与大肠杆菌S17-1作为供体的缀合和可移动质粒作为载体的缀合。最初，使用不相容性组P1的质粒（pRK290和pRK404）。大肠埃希氏菌和绿脓杆菌之间的更有效的穿梭载体pKV1和pKVS1来源于质粒pBR322，并显示出最高的缀合频率（迄今为止在紫色细菌中显示出10（-2）。还证明了根瘤菌可以从载体pHC79中构建的拟南芥基因组粘粒文库中，克隆了编码拟南芥R. viridis的亚基H（puh操纵子），L，M和细胞色素c（puf操纵子）的克隆。分离并鉴定了光合反应中心，为了将两个操纵子连接到基因组上，鉴定了与携带操纵子的克隆重叠的粘粒，无法确定两个操纵子的相对位置，但操纵子必须大于100因此，在绿色假单胞菌中，反应中心的基因组组织与荚膜红细菌的基因组组织不同，为此确定了大约39个碱基对的距离。在对除草剂特丁草胺有抗性的R. viridis中，将puf操纵子克隆到pKVS1中，并通过缀合转移到R. viridis野生型细胞中。所得的超接合剂对除草剂具有抗性，这表明pKVS1结构上的puf操纵子在功能性拟南芥中表达。

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