Amber suppressor tRNAs of phage T5: Gene constructs and suppression efficiency in vivo

N. V. Pan'kova, A. L. Karamyshev, M. G. Shlyapnikov, A. N. Presich, V. N. Ksenzenko

Research output: Contribution to journalArticle

Abstract

The genes of 23 phage T5 amber suppressor tRNAs were constructed using site-directed mutagenesis and were cloned in plasmid pGFIB1 under control of the lipoprotein gene constitutive promoter. Suppression efficiency was measured in vivo after introducing recombinant plasmids into E. coli strains bearing mutations in the lacI gene. Strong amber suppressors (not less than 40% efficiency of the wild type) were obtained with tRNAs specific for amino acids Ala, Gln, Gly, His, Lys, Ser, and Tyr. Suppressor activity was considerably enhanced by additional substitutions which restored pairing in the D stem (tRN AAspCUA) and in the T stem (tRN AGluCUA) outside the anticodon, as well as by substitution U32C in the anticodon loop of tRN AArgCUA .

Original languageEnglish (US)
Pages (from-to)846-853
Number of pages8
JournalMolecular Biology
Volume32
Issue number6
StatePublished - Nov 1 1998

Keywords

  • Amber suppressor tRNA
  • Phage T5
  • Site-directed mutagenesis
  • Suppression efficiency

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology

Fingerprint Dive into the research topics of 'Amber suppressor tRNAs of phage T5: Gene constructs and suppression efficiency in vivo'. Together they form a unique fingerprint.

  • Cite this

    Pan'kova, N. V., Karamyshev, A. L., Shlyapnikov, M. G., Presich, A. N., & Ksenzenko, V. N. (1998). Amber suppressor tRNAs of phage T5: Gene constructs and suppression efficiency in vivo. Molecular Biology, 32(6), 846-853.