Shimotsu H, Kawamura F, Kobayashi Y, Saito H
Proc Natl Acad Sci U S A. 1983 Feb;80(3):658-62. doi: 10.1073/pnas.80.3.658.
We have determined the sequence of a 1,162-base-pair DNA fragment containing a spo0F gene which is required for an early stage of sporulation in Bacillus subtilis. The sequence has only one long open reading frame consisting of 173 codons, which has been confirmed to be the spo0F cistron by DNA-mediated transformation and in vitro transcription. In UV-irradiated "maxicells" containing pUBSF13, the plasmid that carries cloned spo0F gene, we have observed the synthesis of a 20-kilodalton polypeptide that is absent from cells carrying a vector plasmid pUB110. The molecular weight of this protein is in agreement with the calculated molecular weight of the spo0F gene product (Mr, 19,065). The putative promoter sequences of spo0F gene were 5' T-A-T-A-A-T 3' at -10 and 5' T-T-G-A-T-T 3' at -35. An octamer sequence, 5' A-A-A-G-G-A-G-G 3', situated 8 base pairs prior to the initiation codon was found to be perfectly complementary with the 3' end of 16S ribosomal RNA. This result offers additional evidence for the proposal by Rabinowitz's group that an extensive mRNA-rRNA interaction is a requirement for efficient translation by B. subtilis ribosomes.
我们已经确定了一段1162个碱基对的DNA片段的序列,该片段包含枯草芽孢杆菌芽孢形成早期所需的spo0F基因。该序列只有一个由173个密码子组成的长开放阅读框,通过DNA介导的转化和体外转录已证实其为spo0F顺反子。在含有携带克隆spo0F基因的质粒pUBSF13的紫外线照射的“大细胞”中,我们观察到合成了一种20千道尔顿的多肽,而携带载体质粒pUB110的细胞中则没有这种多肽。这种蛋白质的分子量与spo0F基因产物的计算分子量(Mr,19,065)一致。spo0F基因的推定启动子序列在-10处为5'T-A-T-A-A-T 3',在-35处为5'T-T-G-A-T-T 3'。发现在起始密码子之前8个碱基对处的一个八聚体序列5'A-A-A-G-G-A-G-G 3'与16S核糖体RNA的3'末端完全互补。这一结果为拉比诺维茨小组的提议提供了额外证据,即广泛的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。