Growth at low temperature suppresses readthrough of the UGA stop codon during the expression of Bacillus subtilis flgM gene in Escherichia coli.
The efficient production of recombinant proteins in Escherichia coli requires a proper termination of translation to ensure the synthesis of only the desired product. During the recombinant production of Bacillus subtilis flgM in E. coli, we detected an additional polypeptide of molecular mass higher than the expected, corresponding to a product of a translational readthrough of the UGA stop codon. In this paper we show that the readthrough was abolished when the synthesis of the recombinant protein was carried out at 25 degrees C. The possible causes that contribute to reduce the proportion of readthrough protein species against the correct terminated product are discussed.
González
Beatriz
B
Laboratory of Bioreactors, Plant Division, Genetic Engineering and Biotechnology Center, PO Box 6162, CP 10600, La Habana, Cuba. beatriz.gonzalez@cigb.edu.cu
Ceciliani
Fabrizio
F
Galizzi
Alessandro
A
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018388:Codon, Terminator / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods; D009154:Mutation; D014176:Protein Biosynthesis / Q000502:physiology; D011786:Quality Control; D016364:Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013696:Temperature
D001426:Bacterial Proteins; D018388:Codon, Terminator; D011994:Recombinant Proteins; C071266:FlgM protein, Bacteria