A new method for identifying the amino acid attached to a particular RNA in the cell

被引:15
作者
Suzuki, T
Ueda, T
Watanabe, K
机构
[1] UNIV TOKYO,SCH ENGN,DEPT CHEM & BIOCHEM,BUNKYO KU,TOKYO 113,JAPAN
[2] TOKYO INST TECHNOL,FAC BIOSCI & BIOTECHNOL,DEPT BIOL SCI,MIDORI KU,YOKOHAMA,KANAGAWA 227,JAPAN
关键词
aminoacyl-tRNA (in vivo); amino acid; hybridization; N-acetylation; Candida;
D O I
10.1016/0014-5793(96)00107-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [C-14]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The C-14-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.
引用
收藏
页码:195 / 198
页数:4
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