Rabinovich P M, Iomantas Iu V, Khaĭkinson M Ia, Stepanov A I
Mol Biol (Mosk). 1984 Jan-Feb;18(1):189-96.
Plasmid vectors capable for propagation of Bacillus subtilis DNA fragments containing riboflavin genes were constructed. Cloning of rib operon using pUB110 derivatives was performed in recE4 strain by using sequentional rescue of plasmids containing subfragments of the operon. Also, rib operon was cloned on the vectors containing DNA repeats. It was shown that the presence of direct and inverted repeats within plasmids allows to transform B. subtilis cells by monomers of plasmid DNA. Vectors that contained repeated sequences of DNA and ensured efficient cloning of genetic material in B. subtilis recipient cells were constructed. The use of streptococcal plasmid pSM19035 allowed to obtain vectors which were suitable for cloning large DNA fragments (6 MD and even more) in B. subtilis. A model of B. subtilis transformation by various types of plasmid DNA is presented. The model is in agreement with the general conception of chromosomal DNA transformation.
构建了能够传播含有核黄素基因的枯草芽孢杆菌DNA片段的质粒载体。利用pUB110衍生物对rib操纵子进行克隆,在recE4菌株中通过依次拯救含有该操纵子亚片段的质粒来进行。此外,rib操纵子被克隆到含有DNA重复序列的载体上。结果表明,质粒中正向和反向重复序列的存在使得质粒DNA单体能够转化枯草芽孢杆菌细胞。构建了含有DNA重复序列并确保在枯草芽孢杆菌受体细胞中高效克隆遗传物质的载体。使用链球菌质粒pSM19035可获得适合在枯草芽孢杆菌中克隆大DNA片段(6 MD及以上)的载体。提出了不同类型质粒DNA转化枯草芽孢杆菌的模型。该模型与染色体DNA转化的一般概念一致。