Gilchrist A, Smit J
Department of Microbiology, University of British Columbia, Vancouver, Canada.
J Bacteriol. 1991 Jan;173(2):921-5. doi: 10.1128/jb.173.2.921-925.1991.
We performed plasmid electrotransformation of Caulobacter crescentus strains and obtained up to 3 x 10(8) transformants per micrograms of pKT230. The presence and integrity of the paracrystalline protein surface (S) layer influenced electroporation; caulobacters lacking the S layer were electrotransformed 10 times more efficiently than caulobacters possessing the S layers. A procedure yielding 1,500 transformants per micrograms of pKT230 was developed for a marine caulobacter. Electroporation was used in combination with several genetic techniques, including introduction of ligation mixtures, suicide transposon mutagenesis, gene replacement, and plasmid electrotransfer from Escherichia coli to caulobacters.
我们对新月柄杆菌菌株进行了质粒电转化，每微克pKT230最多可获得3×10⁸个转化子。准晶体蛋白表面（S）层的存在和完整性影响电穿孔；缺乏S层的柄杆菌的电转化效率比具有S层的柄杆菌高10倍。针对一种海洋柄杆菌开发了一种每微克pKT230可产生1500个转化子的方法。电穿孔与多种遗传技术结合使用，包括引入连接混合物、自杀转座子诱变、基因替换以及从大肠杆菌到柄杆菌的质粒电转移。
