嗜热绿硫细菌嗜热栖热菌中通过接合作用进行的基因转移。
Genetic transfer by conjugation in the thermophilic green sulfur bacterium Chlorobium tepidum.
作者信息
Wahlund T M, Madigan M T
机构信息
Department of Microbiology, Southern Illinois University, Carbondale 62901-6508, USA.
出版信息
J Bacteriol. 1995 May;177(9):2583-8. doi: 10.1128/jb.177.9.2583-2588.1995.
Abstract
The broad-host-range IncQ group plasmids pDSK519 and pGSS33 were transferred by conjugation from Escherichia coli into the thermophilic green sulfur bacterium Chlorobium tepidum. C. tepidum exconjugants expressed the kanamycin and ampicillin-chloramphenicol resistances encoded by pDSK519 and pGSS33, respectively. Ampicillin resistance was a particularly good marker for selection in C. tepidum. Both pDSK519 and pGSS33 were stably maintained in C. tepidum at temperatures below 42 degrees C and could be transferred between C. tepidum and E. coli without modifications. Conjugation frequencies ranged from 10(-1) to 10(-4) exconjugants per donor cell, and frequencies of 10(-2) to 10(-3) were consistently obtained when ampicillin resistance was used as a selectable marker. Methods for growth of C. tepidum on agar, isolation of plating strains and antibiotic-resistant mutants of wild-type C. tepidum cells, and optimum conditions for conjugation were also investigated.
摘要
广宿主范围的IncQ群质粒pDSK519和pGSS33通过接合作用从大肠杆菌转移至嗜热绿硫细菌嗜热栖热菌中。嗜热栖热菌接合子分别表达了由pDSK519和pGSS33编码的卡那霉素和氨苄青霉素-氯霉素抗性。氨苄青霉素抗性是在嗜热栖热菌中进行选择的一个特别好的标记。pDSK519和pGSS33在42℃以下的温度下都能在嗜热栖热菌中稳定维持，并且可以在嗜热栖热菌和大肠杆菌之间无修饰地转移。接合频率为每个供体细胞产生10^(-1)至10^(-4)个接合子，当使用氨苄青霉素抗性作为选择标记时，始终能获得10^(-2)至10^(-3)的频率。还研究了嗜热栖热菌在琼脂上的生长方法、平板菌株的分离以及野生型嗜热栖热菌细胞的抗生素抗性突变体，以及接合的最佳条件。
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