Shao Z, Dick W A, Behki R M
Centre for Land and Biological Resources Research, Agriculture Canada, Ottawa, Ontario.
Lett Appl Microbiol. 1995 Oct;21(4):261-6. doi: 10.1111/j.1472-765x.1995.tb01056.x.
The genetic studies of metabolically diverse Rhodococcus spp. have been hampered by the lack of a system of introducing exogenous DNA. The authors improved an existing Escherichia coli-Rhodococcus shuttle vector (pMVS301) by removing much of the DNA not needed for replication and adding a multicloning site. This improved vector (pBS305) is 7.9 kb in length. Its ability to transform Rhodococcus was tested using electroporation parameters optimized for introduction of pMVS301 into Rhodococcus. Transformation efficiencies as high as 10(5) cfu micrograms-1 DNA were obtained although efficiencies varied depending on the Rhodococcus strain tested. The improved vector pBS305 offers great utility for genetic studies of Rhodococcus because its small size enables movement of large inserts of DNA into Rhodococcus, it has multicloning sites, contains a highly selective thiostrepton marker, and can be replicated in both E. coli and Rhodococcus.
代谢多样的红球菌属的遗传学研究因缺乏引入外源DNA的系统而受到阻碍。作者通过去除许多复制不需要的DNA并添加多克隆位点，对现有的大肠杆菌-红球菌穿梭载体（pMVS301）进行了改进。这种改进后的载体（pBS305）长度为7.9 kb。使用针对将pMVS301引入红球菌而优化的电穿孔参数测试了其转化红球菌的能力。尽管转化效率因所测试的红球菌菌株而异，但仍获得了高达10(5) cfu微克-1 DNA的转化效率。改进后的载体pBS305为红球菌的遗传学研究提供了很大的实用性，因为其小尺寸能够使大片段DNA插入红球菌，它具有多克隆位点，含有高度选择性的硫链丝菌素标记，并且可以在大肠杆菌和红球菌中复制。
