Amber-suppressive tRNA from bacteriophage T5: construction of genes and determination of the effectiveness of suppression in vivo
Pan'kova, N.V.; Karamyshev, A.L.; Shliapnikov, M.G.; Presich, A.N.; Ksenzenko, V.N.
Molekuliarnaia Biologiia 32(6): 1004-1012
1998
ISSN/ISBN: 0026-8984 PMID: 9929878 Document Number: 496316
Oligonucleotide site-directed mutagenesis was used to construct 23 genes of amber-suppressor tRNAs of bacteriophage T5. Genes were cloned in plasmid pGFIB1 under control of constitutive promoter of lipoprotein gene. Suppression level was determined in vivo after the introduction of recombinant plasmids into Escherichia coli strains carrying mutations in gene lacI. The strongest amber-suppressors were obtained on the basis of tRNAs specific to amino acids Ala, Gln, Gly, His, Lys, Ser, and Tyr. Additional substitutions beyond the anticodon area resulted in a significant increase in suppressor activity.