通过整合载体的单链DNA转化在链霉菌中进行基因破坏和基因替换。
Gene disruption and gene replacement in Streptomyces via single stranded DNA transformation of integration vectors.
作者信息
Hillemann D, Pühler A, Wohlleben W
机构信息
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, FRG.
出版信息
Nucleic Acids Res. 1991 Feb 25;19(4):727-31. doi: 10.1093/nar/19.4.727.
Abstract
For the isolation of single stranded plasmid DNA, various E. coli and E. coli-Streptomyces shuttle plasmids were equipped with the phage f1 replication origin. The transformation of some representative Streptomyces species with plasmid vectors occurred irrespective of whether single or double stranded DNA was used. In contrast, the transformation of Streptomyces was 10 to 100 times more efficient when an integration vector was in the single stranded form as opposed to the double stranded form. Streptomyces viridochromogenes was transformed by single stranded DNA integration vectors in order to replace the pat by the tsr gene and generate mutants unable to synthesize phosphinothricin-tripeptide (PTT).
摘要
为了分离单链质粒DNA，各种大肠杆菌和大肠杆菌 - 链霉菌穿梭质粒都配备了噬菌体f1复制起点。无论使用单链还是双链DNA，一些代表性链霉菌物种用质粒载体进行转化都能发生。相比之下，当整合载体为单链形式而非双链形式时，链霉菌的转化效率要高10到100倍。为了用tsr基因取代pat并产生无法合成草丁膦三肽（PTT）的突变体，用单链DNA整合载体转化了绿色产色链霉菌。
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