INVITRO SUPPRESSION OF AN AMBER MUTATION BY A CHEMICALLY AMINOACYLATED TRANSFER-RNA PREPARED BY RUNOFF TRANSCRIPTION

被引：83

作者：

NOREN, CJ ^{[1
]}

ANTHONYCAHILL, SJ ^{[1
]}

SUICH, DJ ^{[1
]}

NOREN, KA ^{[1
]}

GRIFFITH, MC ^{[1
]}

SCHULTZ, PG ^{[1
]}

机构：

[1] UNIV CALIF BERKELEY,DEPT CHEM,BERKELEY,CA 94720

来源：

NUCLEIC ACIDS RESEARCH | 1990年 / 18卷 / 01期

关键词：

D O I：

10.1093/nar/18.1.83

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：

An amber suppressor tRNA was prepared in vitro by runoff transcription with T7 RNA polymerase. Both fulllength tRNA and truncated tRNA lacking the 3' terminal pCpA from the acceptor stem could be synthesized from the same DNA template. Truncated runoff suppressor tRNA could be enzymatically ligated to phenylalanyl-pCpA to generate aminoacylated fulllength suppressor tRNA (Phe-tRNACUA). Phe-tRNACUA is capable of suppressing an amber (UAG) mutation in vitro with equivalent efficiency as suppressor prepared by anticodon-loop replacement of a naturally-isolated tRNA. The ease of suppressor tRNA preparation using this method, compared to anticodon-loop replacement, greatly facilitates the use of chemically acylated suppressor tRNA's for site-specifically incorporating unnatural amino acids into proteins. © 1990 Oxford University Press.

引用

页码：83 / 88

页数：6

共 24 条

[21] A BACTERIOPHAGE-T7 RNA-POLYMERASE PROMOTER SYSTEM FOR CONTROLLED EXCLUSIVE EXPRESSION OF SPECIFIC GENES [J].