An inducible expression system of histidine-tagged proteins in Streptomyces lividans for one-step purification by Ni2+ affinity chromatography

被引：0

作者：

Enguita, FJ ^{[1
]}

delaFuente, JL ^{[1
]}

Martin, JF ^{[1
]}

Liras, P ^{[1
]}

机构：

[1] UNIV LEON, FAC BIOL, DEPT ECOL GENET & MICROBIOL, AREA MICROBIOL, E-24071 LEON, SPAIN

来源：

FEMS MICROBIOLOGY LETTERS | 1996年 / 137卷 / 2-3期

关键词：

protein purification; affinity chromatography; expression vector; cephamycin biosynthesis genes;

D O I：

暂无

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

An expression and purification cassette containing the aminoglycoside phosphotransferase gene (aph) as selective marker has been constructed in the Escherichia coli vector pULHis2. DNA fragments inserted in the cassette can be easily subcloned in pIJ699 to give vectors for overexpression of genes in Streptomyces and purification of proteins by a one-step procedure. The expression system uses the thiostrepton-inducible promoter tipA for expression and a six histidine coding nucleotide sequence that is fused in frame to the foreign gene inserted in the polylinker. The pULHis2-derived expression vector has been used satisfactorily to express and to purify the P-7 and P-8 proteins of Nocardia lactamdurans which carry our the methoxylation of cephalosporin C to 7-methoxycephalosporin C.

引用

页码：135 / 140

页数：6

共 17 条

[1] PRODUCTION IN ESCHERICHIA-COLI AND ONE-STEP PURIFICATION OF BIFUNCTIONAL HYBRID PROTEINS WHICH BIND MALTOSE - EXPORT OF THE KLENOW POLYMERASE INTO THE PERIPLASMIC SPACE [J].

BEDOUELLE, H ;

DUPLAY, P .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 171 (03) :541-549

[2] A 2-PROTEIN COMPONENT 7-ALPHA-CEPHEM-METHOXYLASE ENCODED BY 2 GENES OF THE CEPHAMYCIN-C CLUSTER CONVERTS CEPHALOSPORIN-C TO 7-METHOXYCEPHALOSPORIN-C [J].

COQUE, JJR ;

ENGUITA, FJ ;

MARTIN, JF ;

LIRAS, P .

JOURNAL OF BACTERIOLOGY, 1995, 177 (08) :2230-2235

[3]

COQUE JJR, 1993, FEMS MICROBIOL LETT, V110, P91, DOI 10.1111/j.1574-6968.1993.tb06300.x

[4]

CORTES J, 1987, J GEN MICROBIOL, V133, P3165

[5]

GUAN C, 1988, GENE, V67, P21

[6] NEW METAL CHELATE ADSORBENT SELECTIVE FOR PROTEINS AND PEPTIDES CONTAINING NEIGHBORING HISTIDINE-RESIDUES [J].

HOCHULI, E ;

DOBELI, H ;

SCHACHER, A .

JOURNAL OF CHROMATOGRAPHY, 1987, 411 :177-184

[7] PURIFICATION OF HIS-TAGGED PROTEINS IN NONDENATURING CONDITIONS SUGGESTS A CONVENIENT METHOD FOR PROTEIN-INTERACTION STUDIES [J].

HOFFMANN, A ;

ROEDER, RG .

NUCLEIC ACIDS RESEARCH, 1991, 19 (22) :6337-6338

[8]

Hopwood D.A., 1985, GENETIC MANIPULATION

[9] DERIVATIVES OF PUC18 THAT HAVE BGLII SITES FLANKING A MODIFIED MULTIPLE CLONING SITE AND THAT RETAIN THE ABILITY TO IDENTIFY RECOMBINANT CLONES BY VISUAL SCREENING OF ESCHERICHIA-COLI COLONIES [J].

JANSSEN, GR ;

BIBB, MJ .

GENE, 1993, 124 (01) :133-134

[10] PLASMID PIJ699, A MULTI-COPY POSITIVE-SELECTION VECTOR FOR STREPTOMYCES [J].

KIESER, T ;

MELTON, RE .

GENE, 1988, 65 (01) :83-91

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