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Cloning, sequencing and expression of subtilisin Carlsberg from Bacillus licheniformis.
The gene encoding subtilisin Carlsberg from Bacillus licheniformis has been isolated by molecular cloning using a mixture of synthetic oligonucleotides. The entire nucleotide sequence of the coding sequence as well as 5' and 3' flanking sequences have been determined. The deduced amino acid sequence reveals an N-terminal signal peptide consisting of 29 residues, a pro-peptide of 76 residues followed by the mature protein comprising 274 residues. The ATG initiator codon is preceded by two putative overlapping ribosomal binding sequences. A palindromic sequence typical for transcription termination is found downstream from the TAA stop codon. Structural comparisons between different known subtilisin genes reveal extensive homology, particularly in the parts coding for the pro-region and the mature protein. Expression studies in Bacillus subtilis show that the cloned fragment produces a functional enzyme when inserted after a B. subtilis promoter.
PMID: 3001653 [PubMed - indexed for MEDLINE]
PMCID: PMC318961
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Cited by 21 PubMed Central articles
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A posttranslationally regulated protease, VheA, is involved in the liberation of juveniles from parental spheroids in Volvox carteri.
Fukada K, Inoue T, Shiraishi H.
Plant Cell. 2006 Oct; 18(10):2554-66. Epub 2006 Oct 6.
[Plant Cell. 2006]
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Ca2+-dependent maturation of subtilisin from a hyperthermophilic archaeon, Thermococcus kodakaraensis: the propeptide is a potent inhibitor of the mature domain but is not required for its folding.
Pulido M, Saito K, Tanaka S, Koga Y, Morikawa M, Takano K, Kanaya S.
Appl Environ Microbiol. 2006 Jun; 72(6):4154-62.
[Appl Environ Microbiol. 2006]
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Cloning and expression of islandisin, a new thermostable subtilisin from Fervidobacterium islandicum, in Escherichia coli.
Gödde C, Sahm K, Brouns SJ, Kluskens LD, van der Oost J, de Vos WM, Antranikian G.
Appl Environ Microbiol. 2005 Jul; 71(7):3951-8.
[Appl Environ Microbiol. 2005]
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