期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1974
卷号:71
期号:9
页码:3635-3639
DOI:10.1073/pnas.71.9.3635
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:The protein product of the regulatory gene araC can be synthesized in a cell-free, protein-synthesizing system programmed with a {lambda}paraC+B DNA template. Hybrid, renatured phage DNA molecules prepared with DNA from phages {lambda}paraC+B and {lambda}paraC3B (araC3 is a nonsense mutation) were used to program the cell-free synthesis of the araC protein. The findings observed lead to the conclusion that the codogenic strand of the araC gene is on the light strand of the phage DNA. The araB gene is on the heavy strand, as determined by DNA{middle dot}RNA hybridization. Thus, with regard to the standard E. coli map, araC is transcribed in a clockwise direction, whereas transcription of the araBAD operon has a counterclockwise orientation. The technique described should allow one to determine the direction of transcription of any gene that can be incorporated into the genome of a specialized transducing phage.
关键词:protein synthesis in vitro ; clockwise transcription ; L-arabinose ; positive control