Nucleotide sequences of transcription and translation initiation regions in Bacillus phage phi 29 early genes.
Nucleotide sequences of transcription and translation initiation regions in Bacillus phage phi 29 early genes.
复制标题
芽孢杆菌噬菌体 phi 29 早期基因转录和翻译起始区的核苷酸序列。
DOI：
10.1016/s0021-9258(19)68307-9
复制
发表时间：
1982
期刊：
影响因子：
--
通讯作者：
J. Rabinowitz
中科院分区：
文献类型：
--
作者：
C. L. Murray;J. Rabinowitz
在源自枯草芽孢杆菌的无细胞转录-翻译系统中，phi 29 DNA指导Mr = 22400、13900和10500三种主要蛋白质的合成。我们已经确定了这些早期蛋白质在phi 29基因组上的编码区域的位置，我们的结果与遗传证据一致，即22.4千道尔顿的蛋白质是反顺子17 （p17）的产物，13.9千道尔顿的蛋白质是反顺子6 （p6）的产物。这两种13.9千道尔顿和10.5千道尔顿的蛋白编码在一种多顺反电子mRNA上，这种mRNA之前被命名为G3b RNA。我们已经确定了phi 29 DNA的HindIII限制性片段（H片段）的核苷酸序列，该片段编码13.9千道尔顿蛋白，并包含两个早期启动子G3a和G3b。核糖体结合位点的核苷酸序列包含一个多嘌呤区，能够在枯草芽孢杆菌16s rRNA的3'端与9个碱基形成非常稳定的复合物。这种强烈的Shine-Dalgarno互补性支持了广泛的mRNA。rRNA相互作用是枯草芽孢杆菌核糖体有效翻译的必要条件(McLaughlin， J. R., Murray， C. L., and Rabinowitz， J. C.（1981）。化学，256,1183 -11291)。这两个启动子的核苷酸序列与大肠杆菌启动子的“共识”序列非常相似。大肠杆菌RNA聚合酶在体外可同时启动这两种启动子的转录，而枯草芽孢杆菌RNA聚合酶在相同条件下仅利用G3b启动子。
phi 29 DNA directs the synthesis of three major proteins of Mr = 22,400, 13,900, and 10,500 in a cell-free transcription-translation system derived from Bacillus subtilis. We have determined the locations of the coding regions for these early proteins on the phi 29 genome, and our results are in agreement with genetic evidence that the 22.4-kilodalton protein is the product of cistron 17 (p17) and the 13.9-kilodalton protein is the product of cistron 6 (p6). The 13.9-kilodalton and 10.5-kilodalton proteins are encoded on a polycistronic mRNA previously designated G3b RNA. We have determined the nucleotide sequence of a HindIII restriction fragment of phi 29 DNA, the H fragment, that encodes the 13.9-kilodalton protein and contains two early promoters, G3a and G3b. The nucleotide sequence of the ribosome binding site contains a polypurine region capable of forming a very stable complex with nine bases on the 3' end of B. subtilis 16 S rRNA. This strong Shine-Dalgarno complementarity supports the hypothesis that an extensive mRNA . rRNA interaction is a requirement for efficient translation by B. subtilis ribosomes (McLaughlin, J. R., Murray, C. L., and Rabinowitz, J. C. (1981) J. Biol. Chem. 256, 11283-11291). The nucleotide sequences of both promoters are very similar to the “consensus” sequence for Escherichia coli promoters. While E. coli RNA polymerase initiates transcription from both of these promoters in vitro, only the G3b promoter is utilized by B. subtilis RNA polymerase under the same conditions.