Gene transfer system for Rhodopseudomonas viridis
Gene transfer system for Rhodopseudomonas viridis
复制标题
绿色红假单胞菌基因转移系统
DOI：
--
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发表时间：
1989
影响因子：
3.2
通讯作者：
D. Oesterhelt
中科院分区：
文献类型：
--
作者：
F. Lang;D. Oesterhelt
以绿色红假单胞菌S17-I为供体，以可动员载体为载体，建立了绿色红假单胞菌基因转移系统。最初，使用不相容组P1(pRK290和pRK404)的质粒。在绿色弧菌和大肠杆菌之间更有效的穿梭载体pKV1和pKVS1来自pBR322，在紫色细菌中显示出最高的接合频率(10(-2))。还证明了紫花苜蓿根瘤菌可以作为供体与绿色根瘤菌接合。从绿色乳杆菌基因组粘粒文库中分离并鉴定了光合作用反应中心H亚单位(PUH操纵子)、L操纵子(M)和细胞色素c(PUF操纵子)基因。为了连接基因组上的两个操纵子，确定了与携带操纵子的克隆重叠的宇宙体。无法确定两个操纵子的相对位置，但两个操纵子的距离必须超过100千碱基对。因此，绿色假单胞菌的反应中心的基因组结构不同于胶囊红杆菌的反应中心的基因组结构，后者的距离约为39kb。从绿色核桃抗除草剂特丁隆的自发突变株出发，将PUF操纵子克隆到pKVS1中，接合转入绿色核盘菌野生型细胞中。由此得到的外接子对除草剂具有抗性，这表明pKVS1结构上的PUF操纵子在绿色根霉中得到了功能性表达。
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.
影响因子：
2.6
作者：
DITTA, G;SCHMIDHAUSER, T;HELINSKI, DR
通讯作者：
HELINSKI, DR
球形红假单胞菌反应中心 L 亚基的一级结构。
DOI：
10.1073/pnas.81.23.7303
发表时间：
1984
影响因子：
11.1
作者：
Williams,JC;Steiner,LA;Feher,G;Simon,MI
通讯作者：
Simon,MI