Gurskiĭ Ia G, Marimont N Iu, Bibilashvili R Sh
Mol Biol (Mosk). 1992 Sep-Oct;26(5):1080-7.
Influence of increased arginine concentrations of tRNA's corresponding to rare codons AGG and AGA was studied in the model system constructed earlier. The model system is a chimeric gene consisting of CAT gene fragment, part of the gene encoding for alpha-domain of beta-galactosidase E. coli and a series of synthetic inserts enriched with codons AGG and AGA. In order to increase the intracellular tRNA concentration the natural gene of AGA-specific tRNA and the artificial gene of AGG-specific tRNA were cloned in plasmid under the control of p15A ori compatible with co1EI ori and used for maintaining the model gene. It was shown that the artificial AGG-specific tRNA gene produces a functionally active tRNA. A steep rise in the synthesis of polypeptide encoded by the model template containing rare codons was demonstrated when the genes of tRNAs recognizing these codons were propagated in the multicopy plasmid. It was shown that AGA-specific tRNA efficiently translates both AGA and AGG codons while AGG-specific tRNA - only AGG codons.
在之前构建的模型系统中,研究了对应于稀有密码子AGG和AGA的tRNA中精氨酸浓度增加的影响。该模型系统是一个嵌合基因,由CAT基因片段、编码大肠杆菌β-半乳糖苷酶α结构域的基因部分以及一系列富含密码子AGG和AGA的合成插入片段组成。为了提高细胞内tRNA浓度,将AGA特异性tRNA的天然基因和AGG特异性tRNA的人工基因克隆到与colE1 ori兼容的p15A ori控制下的质粒中,并用于维持模型基因。结果表明,人工AGG特异性tRNA基因产生功能活性tRNA。当识别这些密码子的tRNA基因在多拷贝质粒中增殖时,含有稀有密码子的模型模板编码的多肽合成急剧增加。结果表明,AGA特异性tRNA能有效翻译AGA和AGG密码子,而AGG特异性tRNA只能翻译AGG密码子。