Molecular cloning and nucleotide sequence determination of the Bacillus stearothermophilus NCA 1503 superoxide dismutase gene and its overexpression in Escherichia coli.
The gene (sod) encoding Bacillus stearothermophilus Mn-superoxide dismutase (MnSOD) has been cloned in Escherichia coli and its entire nucleotide sequence determined. With the exception of the post-translationally cleaved N-terminal methionine residue, the predicted amino acid sequence exhibits complete identity to the previously determined amino acid sequence. The recombinant MnSOD was shown to be functionally active in E. coli both in vitro and in vivo, and was expressed to 49% of the soluble cell protein by coupling its transcription to the E. coli trp promoter. The sequenced region of DNA was also found to encompass a second open reading frame. The putative encoded polypeptide exhibited no significant primary sequence homology to any currently characterised protein.
Molecular Genetics Group, Division of Biotechnology, PHLS Centre for Applied Microbiology and Research, Salisbury, Wiltshire, UK.
J K
JK
Brehm
S P
SP
Chambers
K J
KJ
Brown
T
T
Atkinson
N P
NP
Minton
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics; D008345:Manganese; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D008345:Manganese; D013482:Superoxide Dismutase