Jahn O, Hartmann R K, Erdmann V A
Freie Universität Berlin, Institut für Biochemie, Federal Republic of Germany.
Eur J Biochem. 1991 May 8;197(3):733-40. doi: 10.1111/j.1432-1033.1991.tb15965.x.
The gene region of Thermus aquaticus corresponding to the distal portion of the S10 operon and to the 5'-portion of the Escherichia coli spc operon was cloned, using the E. coli gene for the ribosomal protein L5 as hybridization probe. The gene arrangement was found to be identical to E. coli, i.e. S17, L14, L24, L5, S14, S8 and L6. Stop and start regions of contiguous cistrons overlap, except for the S14-S8 intergenic region, whose size (67 bases) even exceeds the corresponding spacer regions in E. coli and Bacillus subtilis. A G + C content of 94% in third positions of codons was found in the ribosomal protein genes of T. aquaticus analyzed here. The stop codon of gene S17 (the last gene of the S10 operon in E. coli) and the start codon of gene L14 (the first gene of the spc operon in E. coli) overlap in T. aquaticus, thus leaving no space to accommodate an intergenic promoter preceding spc-operon-encoded genes in T. aquaticus. A possible promoter, localized within the S17 coding region, yielded only weak resistance (20 micrograms/ml) to chloramphenicol in E. coli and therefore could be largely excluded as the main promoter for spc-operon-encoded genes. We failed to detect a structure resembling the protein S8 translational repressor site, located at the beginning of the L5 gene in E. coli, in the corresponding region or any other region in the cloned T. aquaticus spc DNA.
以大肠杆菌核糖体蛋白L5的基因作为杂交探针,克隆了嗜热水生栖热菌中与S10操纵子远端部分及大肠杆菌spc操纵子5'部分相对应的基因区域。发现该基因排列与大肠杆菌相同,即S17、L14、L24、L5、S14、S8和L6。除了S14 - S8基因间区域外,相邻顺反子的终止和起始区域相互重叠,该区域大小(67个碱基)甚至超过了大肠杆菌和枯草芽孢杆菌中的相应间隔区。在此分析的嗜热水生栖热菌核糖体蛋白基因中,密码子第三位的G + C含量为94%。在嗜热水生栖热菌中,基因S17(大肠杆菌S10操纵子的最后一个基因)的终止密码子与基因L14(大肠杆菌spc操纵子的第一个基因)的起始密码子重叠,因此在嗜热水生栖热菌中没有空间容纳spc操纵子编码基因之前的基因间启动子。一个可能位于S17编码区域内的启动子,在大肠杆菌中仅产生对氯霉素的弱抗性(20微克/毫升),因此可基本排除其作为spc操纵子编码基因的主要启动子。我们未能在克隆的嗜热水生栖热菌spc DNA的相应区域或任何其他区域检测到类似于位于大肠杆菌L5基因起始处的蛋白质S8翻译阻遏位点的结构。