Effects of GUG and AUG initiation codons on the expression of lacZ in Escherichia coli

We have replaced the ribosomal binding site (RBS) of the lacZ gene of E. coli by those of the maturation (A) gene of phage MS2 and that of the tufA gene. Both RBSs contain a GUG initiation codon. The expression with the tufA RBS is at least 25-fold higher than with the phage RBS. Changing the GUG in...

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Veröffentlicht in:	FEBS letters 1986-03, Vol.197 (1), p.315-320
Hauptverfasser:	Looman, A.C., van Knippenberg, P.H.
Format:	Artikel
Sprache:	eng
Schlagworte:	1 × SSC, 0.15 M NaCl, 0.015 M Na citrate, pH 7.2 Ap, ampicillin Bacteriology Base Sequence beta-Galactosidase - biosynthesis Binding Sites Biological and medical sciences bp, basepair(s) Codon dNTPs, deoxyribose nucleoside 5'-triphosphates ds, doublestranded Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genes, Bacterial Genetics Km, kanamycin Lac Operon Microbiology Mutation nt, nucleotide(s) Peptide Chain Initiation, Translational Protein Biosynthesis R, resistance rATP, ribose adenosine 5 ' -triphosphate RBS, ribosomal binding site rf, replicative form Ribosomes - metabolism RNA, Bacterial - genetics RNA, Messenger - genetics Site-directed mutagenesis tufA gene Phage MS2 Maturation protein Initiation codon Translation efficiency Ribosomal binding site ss, singlestranded
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Zusammenfassung:	We have replaced the ribosomal binding site (RBS) of the lacZ gene of E. coli by those of the maturation (A) gene of phage MS2 and that of the tufA gene. Both RBSs contain a GUG initiation codon. The expression with the tufA RBS is at least 25-fold higher than with the phage RBS. Changing the GUG into AUG results in a 3-fold increase in expression in both cases. In general, higher expression is accompanied by an increase of lac-specific mRNA. It is argued that this is a consequence of the more efficient translation of the mRNA.
ISSN:	0014-5793 1873-3468
DOI:	10.1016/0014-5793(86)80349-0