Construction of Three Hybrid Promoters and Their Properties in Escherichia Coli

Boer, H de; Heyneker, Herbert L.; Comstock, Lisa J.; Wieland, Alice; Vasser, Mark; Horn, Thomas

doi:10.1016/b978-0-12-045560-7.50023-8

Cited by 2 publications

(1 citation statement)

References 21 publications

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“…It has been suggested (Eckhardt and Luhrmann, 1981;Ganoza et al, 1982) (Eckhardt and Luhrmann, 1981;Ganoza et al, 1982 al., 1980). In addition, the Trp leader mRNA has a three base complementarity of unusual composition, yet when fused to the human growth hormone gene (Goeddel et al, 1979), for example, it results in a highly efficient translation system (de Boer et al, 1982). In agreement with these findings is our recent report (de Boer et al, 1983b) which shows that an increase in the complementarity from 4 to 8 or 13 bp results in a 50Wo decrease in expression level.…”

Section: Comparison Of Mutants That Differ Onlj At Position -3supporting

confidence: 70%

Mutagenesis of the three bases preceding the start codon of the beta-galactosidase mRNA and its effect on translation in Escherichia coli.

Hui¹,

Hayflick²,

Dinkelspiel³

et al. 1984

The EMBO Journal

126

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The effect on the translation efficiency of various mutations in the three bases (the -1 triplet) that precede the AUG start codon of the 3-galactosidase mRNA in Escherichia coli was studied. Of the 39 mutants examined, the level of expression varies over a 20-fold range. The most favorable combinations of bases in the -1 triplet are UAU and CUU. The expression levels in the mutants with UUC, UCA or AGG as the -1 triplet are 20-fold lower than those with UAU or CUU. In general, a U residue immediately preceding the start codon is more favorable for expression than any other base; furthermore, an A residue at the -2 position enhances the translation efficiency in most instances. In both cases, however, the degree of enhancement depends on its context, i.e. the neighboring bases. Although the rules derived from this study are complex, the results show that mutations in any of the three bases preceding the start codon can strongly affect the translational efficiency of the 3-galactosidase mRNA.

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