NUCLEOTIDE-SEQUENCE AND EXPRESSION OF THE ALGE GENE INVOLVED IN ALGINATE BIOSYNTHESIS BY PSEUDOMONAS-AERUGINOSA

被引:43
作者
CHU, L [1 ]
MAY, TB [1 ]
CHAKRABARTY, AM [1 ]
MISRA, TK [1 ]
机构
[1] UNIV ILLINOIS,COLL MED,DEPT MICROBIOL & IMMUNOL M-C 790,835 S WOLCOTT AVE,CHICAGO,IL 60612
关键词
RECOMBINANT DNA; EXOPOLYSACCHARIDE; SIGNAL PEPTIDE; EXOPOLYSACCHARIDE BIOSYNTHESIS; INVERTED REPEATS; PROMOTER; LUNGS; CYSTIC FIBROSIS; INFECTION;
D O I
10.1016/0378-1119(91)90290-R
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Alginate (Alg), a random polymer of mannuronic acid and guluronic acid residues, is synthesized and secreted by Pseudomonas aeruginosa primarily during its infection of the lungs of cystic fibrosis patients. The molecular biology and biochemistry of the enzymatic steps leading to the production of the Alg precursor GDP-mannuronic acid have been elucidated, but the mechanism of polymer formation and export of Alg are not understood. We report the nucleotide sequence of a 2.4-kb DNA fragment containing the algE gene, previously designated alg76, encoding the AlgE protein (M(r) 54361) that is believed to be involved in these late steps of Alg biosynthesis. Expression of algE appears to occur from its own promoter. The promoter region contains several direct and inverted repeat sequences and shares structural similarity with promoters of several other alg genes from P. aeruginosa. In addition, the AlgE protein was overproduced from the tac promoter in P. aeruginosa. N-terminal amino acid sequence analysis showed that the polypeptide contains a signal peptide which is cleaved to form the mature protein during AlgE export from the cell cytoplasm.
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页码:1 / 10
页数:10
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