van Wezel G P, Bibb M J
John Innes Centre, Norwich Research Park, Colney, UK.
Gene. 1996 Dec 5;182(1-2):229-30. doi: 10.1016/s0378-1119(96)00563-x.
We describe an Escherichia coli plasmid, pIJ2581, that can be used for the efficient construction of stable gene disruptants and of gene deletions in Streptomyces. Integration of pIJ2581 derivatives carrying chromosomal sequences is achieved by selecting for plasmid-encoded thiostrepton resistance, while plasmid excision is secured by counter-selection of the pIJ2581 glkA gene, which confers sensitivity to 2-deoxyglucose.
我们描述了一种大肠杆菌质粒pIJ2581,它可用于在链霉菌中高效构建稳定的基因破坏体和基因缺失体。携带染色体序列的pIJ2581衍生物通过选择质粒编码的硫链丝菌素抗性来实现整合,而质粒切除则通过对赋予对2-脱氧葡萄糖敏感性的pIJ2581 glkA基因进行反选择来确保。