A strong promoter provided with the gene encoding arginyl-tRNA synthetase(argS) from Escherichia coli

Mo Fang Liu, Tong Li, Zhao Bao Yin, Min Gang Xu, En Duo Wang, Yin Lai Wang

Research output: Contribution to journalArticlepeer-review

Abstract

Previous studies showed that the gene argS encoding the arginyl-tRNA synthetase(ArgRS) from Escherichia coli(E. coli), was overexpressed 1 000-fold in the E. coli transformant TGl/pUC-argS, while the gene leuS, encoding the leucyl-tRNA synthetase(LeuRS) from E. coli, was only overproduced 35-fold in the same case. To investigate why the expression of these two aminoacyl-tRNA synthetase genes is so different, a fused gene (termed parg-leuS) was constructed by replacement of the 5′ flanking region of leuS to 5′ flanking region of argS. In the E. coli transformant TG1/pUC-pargleuS, the activity of LeuRS was only improved 8. 5-fold, which was much lower than that of the transformant harboring the recombinant plasmid pUC18-leuS or pKK-leuS. However, by RNA dot hybridization the amount of mRNA produced in the transcription of parg-leuS was about 5 times that of the wild type leuS, and was similar to that of pKK-leuS, suggesting that the promoter of argS is very strong. Analysis of the secondary structure around the initiation codon among three mRNAs showed that the secondary structure of the mRNA from parg-leuS was the strongest of the three mRNAs. From the results, it could be deduced that expression of the fused gene parg-leuS might be controlled at the translational level and the strong secondary structure of this mRNA may hinder translation initiation and result in a low translation efficiency.

Original languageEnglish (US)
Pages (from-to)435-440
Number of pages6
JournalActa Biochimica et Biophysica Sinica
Volume32
Issue number5
StatePublished - Dec 1 2000
Externally publishedYes

Keywords

  • Arginyl-tRNA synthetase
  • Gene expression
  • Leucyl-tRNA synthetase
  • Strong promoter

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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