The influence of messenger RNA secondary structure on expression of an immunoglobulin heavy chain inEscherichia coli
- 1 January 1984
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 12 (9) , 3937-3950
- https://doi.org/10.1093/nar/12.9.3937
Abstract
A gene for murine mu heavy chain immunoglobulin has been inserted into a bacterial expression plasmid containing the Escherichia coli trp promoter and ribosome binding site. A low level expression of mu protein was detected. Secondary structure analysis showed the presence of a hairpin loop burying the mu initiation codon. Alteration of secondary structure at this site by oligonucleotide replacement mutagenesis revealed a correlation between mu expression levels and accessibility of the ribosome binding site. Abolition of secondary structure increased mu protein expression over ninety-fold, to a level approximately equal to that of a trpE -mu fusion protein using the native trpE ribosome binding site.Keywords
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