PMID: 7512814Mar 1, 1994Paper

Cloning and expression in Escherichia coli of the gene encoding the Proteus vulgaris chondroitin ABC lyase

Applied Microbiology and Biotechnology
N SatoS Kimura

Abstract

The structural gene encoding chondroitin ABC lyase from Proteus vulgaris was cloned and sequenced. This gene consists of a single open reading frame of 3,063 bp, including a sequence (72 bp) for a possible secretory protein leader peptide, preceded by a Shine-Dalgarno ribosomal binding site. Promoter-like and rho-independent terminator sequences were detected upstream and downstream of the open reading frame, respectively. The G+C content of the coding region was 38.6%. The transcription startpoint was located 41-bp upstream of the initiation codon (ATG). Chondroitin ABC lyase is composed of 997 amino acids, and has a relative molecular mass of 112,635. When the 5.2-kb fragment containing the 1.2-kb upstream from the gene was inserted into pSTV29, and cloned in Escherichia coli, chondroitin ABC lyase was induced in the medium containing chondroitin-6-sulfate as the carbon source. On the other hand, when a 4.2-kb fragment containing only 0.2 kb upstream was inserted into pSTV29(pCHS delta 6), and pCHS delta 6 was introduced into E. coli, the enzyme was constitutively produced, even in medium containing glucose as the carbon source. By immunoblot analysis, the polypeptide synthesized by E. coli cells carrying pCHS delta 6 appeare...Continue Reading

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Citations

Oct 14, 2008·The Journal of Biological Chemistry·Vikas PrabhakarRam Sasisekharan
Oct 30, 2016·International Journal of Biological Macromolecules·Sara DaneshjouKhosro Khajeh
Jan 13, 2018·FEMS Microbiology Reviews·Didier Ndeh, Harry J Gilbert
Aug 20, 2005·The Biochemical Journal·Vikas PrabhakarRam Sasisekharan
Feb 26, 2000·FEMS Microbiology Letters·W L HynesL J Aridgides
Feb 17, 2000·FEMS Microbiology Letters·W L Hynes, S L Walton
Dec 2, 2019·International Journal of Biological Macromolecules·Xingyu LuLingtian Wu
Sep 2, 2020·International Journal of Biological Macromolecules·Zhelun ZhangFeng Han
Apr 23, 2003·Journal of Molecular Biology·Weijun HuangMiroslaw Cygler

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