ESCHERICHIA-COLI THYMIDYLATE SYNTHASE - AMINO-ACID SUBSTITUTIONS BY SUPPRESSION OF AMBER NONSENSE MUTATIONS

被引:63
作者
MICHAELS, ML
KIM, CW
MATTHEWS, DA
MILLER, JH
机构
[1] UNIV CALIF LOS ANGELES, DEPT BIOL, LOS ANGELES, CA 90024 USA
[2] AGOURON PHARMACEUT, LA JOLLA, CA 92037 USA
关键词
function; site-directed mutagenesis; structure; Suppressor tRNAs;
D O I
10.1073/pnas.87.10.3957
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
By using site-directed oligonucleotide mutagenesis, amber nonsense stop codons (5'-TAG-3') have been introduced at 20 sites in the Escherichia coli thymidylate synthase gene. By transforming the thyA mutant plasmids into 13 strains, each of which harbor different amber suppressor tRNAs, we were able to generate over 245 amino acid substitutions in E. coli thymidylate synthase (EC 2.1.1.45). Growth characteristics of these mutants have been studied, yielding a body of information that includes some surprising results in light of the recently published crystal structure of the enzyme.
引用
收藏
页码:3957 / 3961
页数:5
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