Cortese R, Melton D, Tranquilla T, Smith J D
Nucleic Acids Res. 1978 Dec;5(12):4593-611.
Transfer RNA genes of the nematode Caenorhabditis elegans have been cloned in E. coli using the plasmid Col E1 as vector. The tRNAs coded by 3 hybrid plasmids were purified by hybridisation of labelled nematode tRNA with the plasmid DNAs. Each plasmid appears to code for a single distinct tRNA species. The expression of the cloned DNAs was analysed in vivo by injection into nuclei of Xenopus laevis oocytes. Evidence is presented which suggests that these nematode tRNA genes are accurately transcribed and processed in frog oocytes. Analysis of one hybrid plasmid shows that a 300 base pair DNA fragment contains both the structural gene and those regions required for its transcription in vivo. The results show that cloned eukaryotic DNAs from a heterologous source can be tested for functional gene activity in X. laevis oocytes.
利用质粒Col E1作为载体，已在大肠杆菌中克隆了线虫秀丽隐杆线虫的转运RNA基因。通过用标记的线虫tRNA与质粒DNA杂交，纯化了由3种杂交质粒编码的tRNA。每个质粒似乎编码一种独特的tRNA。通过注射到非洲爪蟾卵母细胞核中，在体内分析了克隆DNA的表达。有证据表明，这些线虫tRNA基因在蛙卵母细胞中能被准确转录和加工。对一种杂交质粒的分析表明，一个300碱基对的DNA片段既包含结构基因，也包含其在体内转录所需的区域。结果表明，来自异源的克隆真核DNA可在非洲爪蟾卵母细胞中检测其功能基因活性。
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