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应用基因工程技术，设计并化合成了编码大鼠肝异亮氨酸转移RNA(tRNA~(Ⅰle))的基因。大鼠肝tRNA~(Ⅰle)基因由77bp组成。为了便于克隆与转录，在其5＇端添加了BamHⅠ、SfiⅠ限制性内切酶位点及T7启动于序列，3＇端添加了BstNⅠ及HindⅡ酶切位点，基因全长为120bp，被克隆至M13mp18载体上，再用之转染E．coliJM109．用分子杂交、PCR及酶切分析筛选出阳性克隆，序列分析证实与所设计的基因完全一致。大鼠肝tRNA~(Ⅰle)基因合成与克隆的成功，为进一步tRNA~(Ⅰle)功能与构象的研究打下基础。
Genes encoding rat liver isoleucine transfer RNA (tRNA ~ (Ile)) were designed and synthesized by genetic engineering. Rat liver tRNA ~ (Ile) gene consists of 77bp. In order to facilitate cloning and transcription, BamH I, Sfi I restriction endonuclease sites and T7 promoter sequences were added to the 5 ’end. BstNI and Hind II restriction sites were added to the 3’ end. The full length of the gene was 120 bp and cloned To M13mp18 vector, and then transfection E. coliJM109. The positive clones were screened by molecular hybridization, PCR and enzyme digestion analysis. The sequence analysis confirmed that they were identical with the designed genes. The successful synthesis and cloning of rat tRNA ~ (le) gene lay the foundation for the further study of the function and conformation of tRNA ~ (le).