Kozlov Iu V, Shilov A A
Mol Biol (Mosk). 1975 Jan-Feb;9(1):70-7.
The paper deals with the comparative investigation of initiation and in vitro RNA synthesis on DNA template by E. coli RNA polymerase and B-form of calf thymus RNA polymerase. It was shown in hybridization experiments that in the range of Cot values between 10(2) and 10(4) RNA synthesized by calf thymus RNA polymerase was hybridized with homologous DNA more effectively than RNA synthesized by E. coli RNA polymerase. No differences were observed in the case of low Cot values. RNA chains synthesized by calf thymus RNA polymerase contained in average about 300-600 nucleotides per chain as determined in the kinetic experiments with ATP-gamma-32P and GTP-gamma-32P. These values are in average 5-10 times lower than in the case of bacterial enzyme. The data presented show that calf thymus and E. coli RNA polymerases initiate the RNA synthesis at apparently different sites of calf thymus DNA. The results obtained make the possibility of specific transcription of eucaryotic DNA by bacterial RNA polymerase to be doubtful.
本文研究了大肠杆菌RNA聚合酶和小牛胸腺B型RNA聚合酶在DNA模板上起始和体外RNA合成的比较。杂交实验表明,在Cot值为10²至10⁴的范围内,小牛胸腺RNA聚合酶合成的RNA与同源DNA的杂交比大肠杆菌RNA聚合酶合成的RNA更有效。在低Cot值情况下未观察到差异。用ATP-γ-³²P和GTP-γ-³²P进行的动力学实验测定,小牛胸腺RNA聚合酶合成的RNA链平均每条链约含300 - 600个核苷酸。这些值平均比细菌酶的情况低5 - 10倍。所呈现的数据表明,小牛胸腺和大肠杆菌RNA聚合酶在小牛胸腺DNA的明显不同位点起始RNA合成。所得结果使细菌RNA聚合酶对真核DNA进行特异性转录的可能性受到质疑。