The influence of messenger RNA secondary structure on expression of an immunoglobulin heavy chain in Escherichia coli

Abstract

A gene for murine a heavy chain immunoglobulin has been inserted into a bacterial expression plasmid containing the Eacherichia coli trp promoter and ribosome binding site. A low level expression of μ protein was detected. Secondary structure analysis showed the presence of a hairpin loop burying the μ initiation codon. Alteration of secondary structure at this site by oligonucleotide replacement mutagenesis revealed a correlation between μ expression levels and accessibility of the ribosome binding site. Abolition of secondary structure increased μ protein expression over ninety-fold, to a level approximately equal to that of a trpE-μ fusion protein using the native trpE ribosorae binding site. © 1984 IRL Press Limited, Oxford, England.