Goddard J P, Squire M, Bienz M, Smith J D
Nucleic Acids Res. 1983 May 11;11(9):2551-62. doi: 10.1093/nar/11.9.2551.
A mixture of low molecular weight RNAs, in which only tRNAs were radiolabelled, was used as a hybridisation probe to select for tRNA-like sequences within a bank of human genomic DNA in lambda Charon 4A. A restriction enzyme digest of one of the selected lambda Charon 4A recombinants contained two fragments (2.4 Kb & 1.8 Kb) which hybridised tRNA and which, when subcloned into pAT153, were transcribed in Xenopus oocyte nuclei. Analysis of the subcloned 2.4 Kb fragment, which was of remarkably high transcriptional activity, revealed the presence of a single gene for tRNAGlu in the middle of the fragment. The sequence immediately preceding the gene has the potential for forming a tRNA-like structure.
一种仅对tRNA进行放射性标记的低分子量RNA混合物，被用作杂交探针，以在λ噬菌体Charon 4A载体的人类基因组DNA文库中筛选tRNA样序列。对其中一个选定的λ噬菌体Charon 4A重组体进行限制性内切酶消化，得到两个片段（2.4 Kb和1.8 Kb），它们与tRNA杂交，并且当亚克隆到pAT153中时，能在非洲爪蟾卵母细胞核中进行转录。对转录活性非常高的亚克隆2.4 Kb片段进行分析，发现在该片段中间存在一个tRNAGlu单基因。该基因之前的序列有可能形成类似tRNA的结构。
