通过使用反义（互补）寡脱氧核糖核苷酸敲除未分级的大肠杆菌tRNA中的特定tRNA种类。

Knocking out a specific tRNA species within unfractionated Escherichia coli tRNA by using antisense (complementary) oligodeoxyribonucleotides.

作者信息

Kanda T, Takai K, Yokoyama S, Takaku H

机构信息

Department of Industrial Chemistry, Faculty of Engineering, Chiba Institute of Technology, Narashino, Japan.

出版信息

FEBS Lett. 1998 Dec 4;440(3):273-6. doi: 10.1016/s0014-5793(98)01471-9.

DOI:10.1016/s0014-5793(98)01471-9

PMID:9872385

Abstract

Methods for the preparation of an Escherichia coli tRNA mixture lacking one or a few specific tRNA species can be the basis for future applications of cell-free protein synthesis. We demonstrate here that virtually a single tRNA species in a crude E. coli tRNA mixture can be knocked out by an antisense (complementary) oligodeoxyribonucleotide. One out of five oligomers complementary to tRNA(Asp) blocked the aspartylation almost completely, while minimally affecting the aminoacylation with other 13 amino acids tested. This 'knockout' tRNA behaved similarly to the untreated tRNA in a cell-free translation of an mRNA lacking Asp codons.

摘要

制备缺少一种或几种特定tRNA种类的大肠杆菌tRNA混合物的方法，可作为无细胞蛋白质合成未来应用的基础。我们在此证明，在大肠杆菌粗制tRNA混合物中，几乎单一的tRNA种类可被反义（互补）寡脱氧核糖核苷酸敲除。与tRNA(Asp)互补的五种寡聚物中的一种几乎完全阻断了天冬酰胺化，同时对所测试的其他13种氨基酸的氨酰化影响极小。这种“敲除”的tRNA在缺乏天冬酰胺密码子的mRNA的无细胞翻译中，表现得与未处理的tRNA相似。