SEQUENCE OF THE ALGL GENE OF PSEUDOMONAS-AERUGINOSA AND PURIFICATION OF ITS ALGINATE LYASE PRODUCT

被引：72

作者：

BOYD, A ^{[1
]}

GHOSH, M ^{[1
]}

MAY, TB ^{[1
]}

SHINABARGER, D ^{[1
]}

KEOGH, R ^{[1
]}

CHAKRABARTY, AM ^{[1
]}

机构：

[1] UNIV ILLINOIS,COLL MED,DEPT MICROBIOL & IMMUNOL,MC 790,835 S WOLCOTT AVE,CHICAGO,IL 60612

来源：

GENE | 1993年 / 131卷 / 01期

关键词：

RECOMBINANT DNA; EXOPOLYSACCHARIDE; ALGINATE DEGRADATION; INSERTION MUTAGENESIS; SOUTHERN HYBRIDIZATION; POLYMANNURONATE;

D O I：

10.1016/0378-1119(93)90662-M

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The alginate lyase-encoding gene (algL) of Pseudomonas aeruginosa was localized to a 1.7-kb EcoRI-XbaI fragment within the alginate biosynthetic gene cluster at 34 minutes on the chromosome. The nucleotide sequence of this DNA fragment revealed an ORF encoding a protein of M(r) 40 885 which is transcribed in the same orientation as the other alg genes within the biosynthetic gene cluster. The predicted protein has a potential N-terminal signal peptide which is consistent with its proposed periplasmic location. The AlgL protein was overproduced in Escherichia coli and purified. The purified protein was shown to have alginate lyase activity. In addition, an algL insertion mutant of the mucoid P. aeruginosa 8830 was constructed. This mutant (alm 1) had a nonmucoid phenotype due to a polar effect on the transcription of an essential alg gene, algA. Thus, the algL gene is located within a region of the alginate biosynthetic gene cluster that appears to be non-essential for alginate production.

引用

页码：1 / 8

页数：8

共 29 条

[1] ACTIVITY OF THE HYBRID TRP-LAC(TAC) PROMOTER OF ESCHERICHIA-COLI IN PSEUDOMONAS-PUTIDA - CONSTRUCTION OF BROAD-HOST-RANGE, CONTROLLED-EXPRESSION VECTORS [J].

BAGDASARIAN, MM ;

AMANN, E ;

LURZ, R ;

RUCKERT, B ;

BAGDASARIAN, M .

GENE, 1983, 26 (2-3) :273-282

[2] CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM [J].

BOLIVAR, F ;

RODRIGUEZ, RL ;

GREENE, PJ ;

BETLACH, MC ;

HEYNEKER, HL ;

BOYER, HW ;

CROSA, JH ;

FALKOW, S .

GENE, 1977, 2 (02) :95-113

[3] CLONING OF PSEUDOMONAS-AERUGINOSA ALGG, WHICH CONTROLS ALGINATE STRUCTURE [J].

CHITNIS, CE ;

OHMAN, DE .

JOURNAL OF BACTERIOLOGY, 1990, 172 (06) :2894-2900

[4] CLONING OF GENES-CONTROLLING ALGINATE BIOSYNTHESIS FROM A MUCOID CYSTIC-FIBROSIS ISOLATE OF PSEUDOMONAS-AERUGINOSA [J].

DARZINS, A ;

CHAKRABARTY, AM .

JOURNAL OF BACTERIOLOGY, 1984, 159 (01) :9-18

[5] NUCLEOTIDE-SEQUENCE ANALYSIS OF THE PHOSPHOMANNOSE ISOMERASE GENE (PMI) OF PSEUDOMONAS-AERUGINOSA AND COMPARISON WITH THE CORRESPONDING ESCHERICHIA-COLI GENE MANA [J].

DARZINS, A ;

FRANTZ, B ;

VANAGS, RI ;

CHAKRABARTY, AM .

GENE, 1986, 42 (03) :293-302

[6] LOCALIZATION OF O-ACETYL GROUPS OF BACTERIAL ALGINATE [J].

DAVIDSON, IW ;

SUTHERLAND, IW ;

LAWSON, CJ .

JOURNAL OF GENERAL MICROBIOLOGY, 1977, 98 (FEB) :603-606

[7] PARTIAL-PURIFICATION AND CHARACTERIZATION OF A POLYMANNURONIC ACID DEPOLYMERASE PRODUCED BY A MUCOID STRAIN OF PSEUDOMONAS-AERUGINOSA ISOLATED FROM A PATIENT WITH CYSTIC-FIBROSIS [J].

DUNNE, WM ;

BUCKMIRE, FLA .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1985, 50 (03) :562-567

[8]

EFTEKHAR F, 1991, 91ST GEN M AM SOC MI

[9] PRODUCTION AND CHARACTERIZATION OF SLIME POLYSACCHARIDE OF PSEUDOMONAS-AERUGINOSA [J].

EVANS, LR ;

LINKER, A .

JOURNAL OF BACTERIOLOGY, 1973, 116 (02) :915-924

[10] REPLICATION OF AN ORIGIN-CONTAINING DERIVATIVE OF PLASMID RK2 DEPENDENT ON A PLASMID FUNCTION PROVIDED IN TRANS [J].

FIGURSKI, DH ;

HELINSKI, DR .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (04) :1648-1652

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