Shortridge R D, Pirtle I L, Pirtle R M
Gene. 1985;33(3):269-77. doi: 10.1016/0378-1119(85)90234-3.
A phage lambda clone containing a 13.1-kb human DNA fragment was isolated and found to contain a tRNA gene encoding a glycine tRNA. The nucleotide sequence of the gene and its flanking regions has been determined. The gene does not have an intervening sequence nor does it encode the 3'-terminal CCA sequence found in mature tRNAs. Although this tRNA gene has an anticodon sequence of CCC, it has a striking homology (96%) with a human glycine tRNA which has an anticodon of GCC. As in other eukaryotic tRNA genes, the coding region contains a characteristic internal split promoter sequence, and the 3'-flanking region has a typical RNA polymerase III termination site of five consecutive T residues. There is no apparent sequence in the 5'-flanking region which could serve as a regulatory element. This gene is accurately transcribed in vitro by RNA polymerase III using a HeLa cell-free system. During the course of in vitro transcription, larger precursor tRNAGlyCCC transcripts are processed to yield a mature-sized tRNA product. A precursor-product relationship was established by comparing the ribonuclease A fingerprints of the precursor and product tRNA transcripts.
分离出一个含有13.1 kb人类DNA片段的λ噬菌体克隆，发现其含有一个编码甘氨酸tRNA的tRNA基因。已确定该基因及其侧翼区域的核苷酸序列。该基因没有内含序列，也不编码成熟tRNA中存在的3'-末端CCA序列。尽管这个tRNA基因的反密码子序列为CCC，但它与反密码子为GCC的人类甘氨酸tRNA具有显著的同源性（96%）。与其他真核tRNA基因一样，编码区含有特征性的内部分裂启动子序列，3'-侧翼区域有一个由五个连续T残基组成的典型RNA聚合酶III终止位点。5'-侧翼区域没有明显可作为调控元件的序列。使用HeLa无细胞系统，该基因在体外能被RNA聚合酶III准确转录。在体外转录过程中，较大的前体tRNAGlyCCC转录本被加工产生成熟大小的tRNA产物。通过比较前体和产物tRNA转录本的核糖核酸酶A指纹图谱，建立了前体-产物关系。
