Selvaraj G, Iyer V N
Plasmid. 1985 Jan;13(1):70-4. doi: 10.1016/0147-619x(85)90057-5.
A mobilizable cosmid derivative of an IncP group plasmid was constructed by cloning the oriT region of RK2, a wide host-range plasmid, and the minimal DNA sequence of bacteriophage lambda required for efficient packaging in vitro. This cosmid is 13 kb in size and has unique restriction sites for EcoRI, XhoI, HindIII, and SalI. The XhoI and HindIII sites are within the kanamycin-resistance gene and the SalI site is in the tetracycline-resistance gene. This plasmid was mobilizable from an Escherichia coli donor to a number of diverse gram-negative bacteria at a frequency of 0.8 to 10 per 100 donors. This vector is one of the smallest of all wide host-range cosmids described in the literature. As part of this study, another mobilizable IncP group plasmid vector has also been constructed which, in addition to the sites listed above, has a unique BglII site, but which lacks the packager sequence.
通过克隆广宿主范围质粒RK2的oriT区域和体外高效包装所需的噬菌体λ最小DNA序列，构建了IncP组质粒的一种可移动黏粒衍生物。该黏粒大小为13 kb，具有EcoRI、XhoI、HindIII和SalI的独特限制性酶切位点。XhoI和HindIII位点位于卡那霉素抗性基因内，SalI位点位于四环素抗性基因内。该质粒可从大肠杆菌供体以每100个供体0.8至10的频率转移到多种不同的革兰氏阴性细菌中。该载体是文献中描述的所有广宿主范围黏粒中最小的之一。作为本研究的一部分，还构建了另一种可移动的IncP组质粒载体，除上述位点外，它具有独特的BglII位点，但缺少包装序列。
