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

FEBS Letters

Takai, Kazuyuki¹  Takaku, Hiroshi¹  Yokoyama, Shigeyuki²  Kanda, Takayuki¹ 

[1] Department of Industrial Chemistry, Faculty of Engineering, Chiba Institute of Technology, 2-17-1, Tsudanuma, Narashino, Chiba 275-0016, Japan;Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

关键词: Cell-free protein synthesis;    Antisense oligodeoxyribonucleotide;    tRNA;    Aminoacylation;    Escherichia coli;   

DOI  :  10.1016/S0014-5793(98)01471-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown