Green C J, Vold B S
SRI International, Menlo Park, California 94025.
J Bacteriol. 1993 Aug;175(16):5091-6. doi: 10.1128/jb.175.16.5091-5096.1993.
The polymerase chain reaction (PCR) was used to detect large tRNA gene clusters in Bacillus subtilis, Bacillus badius, Bacillus megaterium, Lactobacillus brevis, Lactobacillus casei, and Staphylococcus aureus. The primers were based on conserved sequences of known gram-positive bacterial tRNA(Arg) and tRNA(Phe) genes. This PCR procedure detected an unusually large tRNA gene cluster in S. aureus. PCR-generated probes were used to identify a 4.5-kb EcoRI fragment that contained 27 tRNA genes immediately 3' to an rRNA operon. Some of these 27 tRNA genes are very similar, but only 1 is exactly repeated in the cluster. The 5' end of this cluster has a gene order similar to that found in the 9- and 21-tRNA gene clusters of B. subtilis. The 3' end of this S. aureus cluster exhibits more similarity to the 16-tRNA gene cluster of B. subtilis. The 24th, 25th, and 26th tRNA genes of this S. aureus tRNA gene cluster code for three similar, unusual Gly-tRNAs that may be used in the synthesis of the peptidoglycan in the cell wall but not in protein synthesis. Southern analysis of restriction digests of S. aureus DNA indicate that there are five to six rRNA operons in this bacterium's genome and that most or all may have large tRNA gene clusters at the 3' end.
采用聚合酶链反应(PCR)检测枯草芽孢杆菌、短芽孢杆菌、巨大芽孢杆菌、短乳杆菌、干酪乳杆菌和金黄色葡萄球菌中的大tRNA基因簇。引物基于已知革兰氏阳性细菌tRNA(Arg)和tRNA(Phe)基因的保守序列。该PCR程序检测到金黄色葡萄球菌中存在一个异常大的tRNA基因簇。PCR产生的探针用于鉴定一个4.5 kb的EcoRI片段,该片段在一个rRNA操纵子的3'端紧邻包含27个tRNA基因。这27个tRNA基因中的一些非常相似,但在该基因簇中只有1个是完全重复的。该基因簇的5'端基因顺序与枯草芽孢杆菌9个和21个tRNA基因簇中的相似。金黄色葡萄球菌这个基因簇的3'端与枯草芽孢杆菌16个tRNA基因簇表现出更多相似性。金黄色葡萄球菌这个tRNA基因簇的第24、25和26个tRNA基因编码三种相似的、不寻常的甘氨酸tRNA,它们可能用于细胞壁中肽聚糖的合成,但不用于蛋白质合成。对金黄色葡萄球菌DNA限制性消化产物的Southern分析表明,该细菌基因组中有五到六个rRNA操纵子,并且大多数或所有操纵子在3'端可能都有大的tRNA基因簇。