Efficiency of a novel non-Shine-Dalgarno and a Shine-Dalgarno consensus sequence to initiate translation in Escherichia coli of genes with different downstream box composition

doi:10.1016/S0944-5013(99)80032-9

Microbiological Research

Volume 154, Issue 1, May 1999, Pages 35-41

https://doi.org/10.1016/S0944-5013(99)80032-9 Get rights and content

Abstract

The efficiency of a novel non-Shine-Dalgarno translational initiator (ACCUACUCGAGUUAG, denoted PL) to promote translation in Escherichia coli was compared with that of the Shine-Dalgarno (SD) consensus sequence (AAGGAGGU) using four reporter genes. The obtained results showed that the genes of pokeweed antiviral protein (PAP I) and human calcitonin (CT) were poorly expressed under the conventional SD and were better expressed under the PL sequence. On the contrary, the genes of human interferon gamma (hIFNγ) and chloramphenicol acetyltransferase (CAT) were highly expressed under SD and poorly expressed under the PL sequence. Computer search revealed a great diversity between the four reporter genes in respect to their complementarity to E. coli 16S rRNA. PAP I and CT genes were rich in nucleotides matching 16S rRNA (called downstream boxes) whereas the complementary domains in the other two (hIFN-γ and CAT) genes were much shorter. The different behavior of the four reporter genes when placed under the translational control of SD and PL sequences was explained by the different binding energy of their mRNAs to the 30S ribosomal subunit.

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Microbiological Research

Abstract

References (31)

The influence of adenine rich motifs in the 3' portion of the ribosome binding site of human IFN-γ gene expression in Escherichia coli

J. Mol. Biol.

Bacteriophage P2 late promoters. II. Comparison of the four late promoter sequences

J. Mol. Biol.

Chemical synthesis and expression of the human calcitonin gene

Gene

Chemical synthesis and expression in E. coli of a human Val₈-calcitonin gene by fusion to a synthetic human interferon-gamma gene

FEBS Lett.

A second putative mRNA binding site on the Escherichia coli ribosome

Gene

Expression, purification and operator binding of the transposon Tn 1721-encoded Tet repressor

J. Mol. Biol.

Traditional control of prokaryotic gene expression

Trends Genet.

Prokaryotic translation: the interactive pathway leading to initiation

Trends Genet.

DNA hybridization electron microscopy: tertiary structure of 16S rRNA

J. Mol. Biol.

Translation of the prophage lambda cl transcript

Cell

The low expression level of pokeweed antiviral protein (PAP) gene in Escherichia coli by the inducible lac promoter is due to inefficient transcription and translation and not to the toxicity of PAP

Archives Biochem. Biophys.

Expression in E. coli K12 strains of two synthetic human calcitonin genes differing in codon composition

Curr. Microbiol.

Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9

Nature

A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding

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Eur. J. Biochem.

Cited by (0)