Denisova L Ia, Zagrebel'nyĭ S N, Zakabunin A I, Kileva E V, Pustoshilova N M
Mol Biol (Mosk). 1985 Mar-Apr;19(2):537-44.
The plasmid pBR322 transcription in the isolated E. coli DNA-dependent RNA-polymerase system was studied. Transcription regions as well as transcripts orientation were defined using both the technique of "criss-cross" hybridization and the annealing of RNA-products with L- and H-strands of plasmid. Summing up obtained data together with available data on promoter localization a circular transcription map of plasmid pBR322 was constructed. Effects of heparin, ion strength and E. coli S-30 system on in vitro transcripts were also studied. The 110-long RNA transcript synthesized in Ori region of pBR322 was found to be the most sensitive to all these factors. RNA-transcripts obtained in in vitro system are able to direct protein synthesis in cell-free S-30 system.
研究了在分离的大肠杆菌DNA依赖性RNA聚合酶系统中质粒pBR322的转录情况。利用“交叉”杂交技术以及RNA产物与质粒L链和H链的退火，确定了转录区域以及转录本的方向。综合所获得的数据以及关于启动子定位的现有数据，构建了质粒pBR322的环状转录图谱。还研究了肝素、离子强度和大肠杆菌S-30系统对体外转录本的影响。发现pBR322 Ori区域合成的110个核苷酸长的RNA转录本对所有这些因素最为敏感。体外系统中获得的RNA转录本能够在无细胞S-30系统中指导蛋白质合成。
