High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes
High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes
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DOI：
10.1016/s0378-1097(97)00392-3
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发表时间：
1997-10-15
影响因子：
2.1
通讯作者：
Smith, CP
中科院分区：
文献类型：
--
作者：
Flett, F;Mersinias, V;Smith, CP
许多链霉菌，包括S. coelicolor A3（2）具有有效的甲基特异性限制系统，该系统可以对异源DNA的引入提供有效的屏障。我们比较了不同类型质粒对S. coelicolor和S. lividans 66，用两株E. coli供体：标准的甲基化熟练菌株S17-1和甲基化缺陷供体ET 12567（pUB 307）。我们证明甲基化缺陷供体可以产生> 10（4）倍的S。与标准供体相比，Coelicolor接合子。在pSET 152衍生物的情况下，其通过位点特异性重组整合到宿主染色体中，接合混合物中高达10%的链霉菌孢子继承质粒。接合过程是足够有效的获得exconjugants与'自杀'交付质粒，因此提供了一个简单的途径进行基因破坏甲基DNA限制性链霉菌，并可能其他细菌。
Many streptomycetes, including S. coelicolor A3(2), possess a potent methyl-specific restriction system which can present an effective barrier to the introduction of heterologous DNA. We have compared the efficiency of intergeneric conjugal transfer of different types of plasmids to S. coelicolor and S. lividans 66 using two E. coli donors: the standard, methylation proficient strain S17-1, and the methylation deficient donor, ET12567(pUB307). We demonstrate that the methylation deficient donor can yield > 10(4)-fold more S. coelicolor exconjugants than the standard donor. In the case of pSET152 derivatives, which integrate into the host chromosome by site-specific recombination, up to 10% of streptomycete spores in the conjugation mixture inherit the plasmid. The conjugation procedure is efficient enough to obtain exconjugants with 'suicide' delivery plasmids and therefore provides a simple route for conducting gene disruptions in methyl DNA-restricting streptomycetes, and possibly other bacteria.