Enhanced readthrough of opal (UGA) stop codons and production of Mycoplasma pneumoniae P1 epitopes in Escherichia coli.
Expression of mycoplasma sequences in Escherichia coli is often hindered by an unusual mycoplasmal codon usage pattern: the UGA stop codon is utilized for tryptophan. This may result in the truncation of cloned proteins and may prevent the detection of products of many cloned genes. To circumvent this translation barrier, we have developed an expression system for the production of mycoplasma proteins in E. coli. The efficiency of an opal suppressor tRNA (trpT176) was augmented with other suppressor mutations (prfB3 or rrsB(SuUGA-delta C1054)) which influence termination events. System efficacy was analyzed by employing suppressor mutations in the expression of TGA-containing sequences from the P1 protein-encoding gene of Mycoplasma pneumoniae.
Smiley
B K
BK
Department of Microbiology, Immunology and Preventive Medicine, Iowa State University, Ames 50011.
Minion
F C
FC
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D017931:DNA Primers; D000939:Epitopes / Q000235:genetics*; D004926:Escherichia coli; D008969:Molecular Sequence Data; D009154:Mutation; D009177:Mycoplasma pneumoniae / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D012364:RNA, Transfer, Trp / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013728:Terminator Regions, Genetic; D014364:Tryptophan / Q000235:genetics
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D003062:Codon; D017931:DNA Primers; D000939:Epitopes; D012364:RNA, Transfer, Trp; D011994:Recombinant Proteins; C035471:adhesin, Mycoplasma pneumoniae; D014364:Tryptophan