title: The influence of messenger RNA secondary structure on expression of an immunoglobulin heavy chain in Escherichia coli.
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.
mesh_terms: D000818:Animals; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D051379:Mice; D009690:Nucleic Acid Conformation; D009876:Operon; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism
chemical_list: D007143:Immunoglobulin Heavy Chains; D007148:Immunoglobulin mu-Chains; D012333:RNA, Messenger; D004262:DNA Restriction Enzymes