Härtlein M, Madern D
European Molecular Biology Laboratory, Grenoble, France.
Nucleic Acids Res. 1987 Dec 23;15(24):10199-210. doi: 10.1093/nar/15.24.10199.
The gene for Escherichia coli leucyl-tRNA synthetase leuS has been cloned by complementation of a leuS temperature sensitive mutant KL231 with an E.coli gene bank DNA. The resulting clones overexpress leucyl-tRNA synthetase (LeuRS) by a factor greater than 50. The DNA sequence of the complete coding regions was determined. The derived N-terminal protein sequence of LeuRS was confirmed by independent protein sequencing of the first 8 aminoacids. Sequence comparison of the LeuRS sequence with all aminoacyl-tRNA synthetase sequences available reveal a significant homology with the valyl-, isoleucyl- and methionyl-enzyme indicating that the genes of these enzymes could have derived from a common ancestor. Sequence comparison with the gene product of the yeast nuclear NAM2-1 suppressor allele curing mitochondrial RNA maturation deficiency reveals about 30% homology.
通过用大肠杆菌基因文库DNA对亮氨酰 - tRNA合成酶leuS的温度敏感突变体KL231进行互补，克隆出了大肠杆菌亮氨酰 - tRNA合成酶leuS的基因。所得克隆使亮氨酰 - tRNA合成酶（LeuRS）的表达量超量50倍以上。测定了完整编码区的DNA序列。通过对前8个氨基酸进行独立的蛋白质测序，证实了推导的LeuRS的N端蛋白质序列。将LeuRS序列与所有可得的氨酰 - tRNA合成酶序列进行比较，发现与缬氨酰、异亮氨酰和甲硫氨酰酶有显著同源性，表明这些酶的基因可能源自一个共同祖先。与酵母核NAM2 - 1抑制等位基因的基因产物进行序列比较，该等位基因可治愈线粒体RNA成熟缺陷，结果显示约有30%的同源性。
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