Lyutzkanova D, Stoilova-Disheva M, Peltekova V
Institute of Microbiology, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria.
Folia Microbiol (Praha). 2001;46(2):119-22. doi: 10.1007/BF02873588.
To clone bifunctional vectors in streptomycetes, it was necessary to define the restriction-modification system of Streptomyces flavopersicus. Plasmid DNA from bifunctional vectors pIJ699 and pXED3-13, isolated from E. coli strains with different methylation systems: E. coli DH5 alpha (dam+ dcm+), E. coli MB5386 (dam dcm), E. coli CB51 (dam dcm+), E. coli NM544 (dam+ dcm), was used for transformation of protoplasts from strain S. flavopersicus. Restriction of dcm-methylated DNA from S. flavopersicus was established. As a host in the intermediate cloning strain E. coli NM544 (dam+ dcm) should be used, as the dcm-transmethylase-dependent strain S. flavopersicus does not process DNA from this strain.
为了在链霉菌中克隆双功能载体,有必要确定黄色链霉菌的限制-修饰系统。从具有不同甲基化系统的大肠杆菌菌株中分离出的双功能载体pIJ699和pXED3-13的质粒DNA:大肠杆菌DH5α(dam+ dcm+)、大肠杆菌MB5386(dam dcm)、大肠杆菌CB51(dam dcm+)、大肠杆菌NM544(dam+ dcm),用于转化黄色链霉菌菌株的原生质体。确定了黄色链霉菌对dcm甲基化DNA的限制作用。作为中间克隆菌株,应使用大肠杆菌NM544(dam+ dcm),因为依赖dcm-转甲基酶的黄色链霉菌菌株不能处理来自该菌株的DNA。