| Title | E. coli tRNA^phe 유전자의 32-T와 39-T 염기의 부위 특이적 돌연변이 | ||
| Author | 김익영 · 이세영 | ||
| Address | 고려대학교 농대 농화학과 | ||
| Bibliography | Korean Journal of Microbiology, 27(3),176-180, 1989 | ||
| DOI | |||
| Key Words | Pseudouridine, Site-specific mutagenesis, tRNA^phe, pheW | ||
| Abstract | There are three pseudourdine (Ψ)bases in the E. coli tRNA^phe. In order to study the function of the pseudouridine bases in the tRNA^phe, changes of bases tRNA^phe gene to other bases were undertaken by the site-specific mutagenesis. Site-specific mutagenesis of T in the pheW gene, a tRNA^phe gene of E. coli, corresponding to the baseat the No.32 position to C and also T corresponding to the base at the No.39 position to C were performed using Kunkel's uracil-containing template method. Identification of mutants were undertaken by the KNA sequencing techniques of the mutated pheW genes and activities of the mutated pheW genes complementing to E. coli NP37 mutant(pheS^-ts) using the recombinant plasmid containing the mutated genes. Neither NP37 harboring pheW gene mutated at No.32 position nor NP37 harboring pheW gene mutated at No.39 position can be grown at non-permissive temperature. The result means that both mutated pheW genes can not complement to E. coli NP37, and that the pseudouridine bases are essential to the activity of the E. coli tRNA^phe in vivo. | ||
| Download PDF | Kor_270302_176-180p.pdf | ||