Blanco G, Rodicio M R, Puglia A M, Méndez C, Thompson C J, Salas J A
Departamento de Biología Funcional, Universidad de Oviedo, Spain.
Mol Microbiol. 1994 May;12(3):375-85. doi: 10.1111/j.1365-2958.1994.tb01027.x.
Changes in expression of ribosomal protein genes during growth and stationary phase of Streptomyces coelicolor A3(2) in liquid medium were studied. Proteins being synthesized were pulse-labelled with [35S]-methionine, separated by two-dimensional polyacrylamide gel electrophoresis, and quantified using the BioImage computer software. Most of the ribosomal proteins were synthesized throughout the life cycle. Exceptions were two proteins whose synthesis drastically decreased at the approach of stationary phase. These two proteins were identified in purified ribosomes as homologues of Escherichia coli ribosomal proteins L10 and L7/L12, using antibodies raised against fusion proteins between these ribosomal proteins and Escherichia coli beta-galactosidase. The genes (rplJ and rplL) encoding the L10 and L7/L12 proteins were contained in a 1.2 kb BamHI fragment that was cloned and sequenced. The linkage and order of the genes coincide with other L10-L7/L12 operons. However, L11 and L1 genes were not present immediately upstream of the L10 gene, as is the case for E. coli and other bacteria. Instead, two open reading frames of unknown function were found immediately upstream of the L10 gene, in an adjacent 1.9 kb BamHI fragment.
研究了天蓝色链霉菌A3(2)在液体培养基中生长和稳定期核糖体蛋白基因的表达变化。用[35S]-甲硫氨酸对正在合成的蛋白质进行脉冲标记,通过二维聚丙烯酰胺凝胶电泳分离,并使用BioImage计算机软件进行定量。大多数核糖体蛋白在整个生命周期中都有合成。例外的是两种蛋白质,它们在稳定期临近时合成急剧减少。利用针对这些核糖体蛋白与大肠杆菌β-半乳糖苷酶之间的融合蛋白产生的抗体,在纯化的核糖体中鉴定出这两种蛋白质是大肠杆菌核糖体蛋白L10和L7/L12的同源物。编码L10和L7/L12蛋白的基因(rplJ和rplL)包含在一个1.2 kb的BamHI片段中,该片段被克隆并测序。这些基因的连锁和顺序与其他L10-L7/L12操纵子一致。然而,与大肠杆菌和其他细菌不同,L11和L1基因并不紧邻L10基因的上游。相反,在紧邻的一个1.9 kb BamHI片段中,在L10基因的紧邻上游发现了两个功能未知的开放阅读框。