In vivo conjugation-coupled recombinational cloning of a Streptomyces lividans chromosomal telomeric DNA using a linear plasmid.
The high efficiency of homologous recombination in yeast and bacteria makes it useful for recombinational cloning of large genomic segments in vivo. The low efficiency of homologous recombination in Streptomyces has hindered the development of this cloning method. Unlike the inefficient mobilization of chromosomal markers, conjugative plasmid transfer is very efficient in Streptomyces. Here we report that the conjugation-coupled recombination procedure can be used to transfer a 10 kb chromosomal telomeric segment of Streptomyces lividans into a linear plasmid. The plasmid predominated in the population of cells after transfer into recipients. These results may promote the development of the recombinational cloning of large chromosomal segments in Streptomyces in vivo.
Tian
Yongqiang
Y
Key Laboratory of Synthetic Biology, Shanghai Institute of Plant Physiology and Ecology, The Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China.
Jiang
Weihong
W
Zhao
Guoping
G
Qin
Zhongjun
Z
D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular / Q000379:methods*; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic; D048170:Streptomyces lividans / Q000235:genetics*; D016615:Telomere
D004269:DNA, Bacterial