CLONING AND SEQUENCING OF A CLUSTER OF GENES ENCODING BRANCHED-CHAIN ALPHA-KETO ACID DEHYDROGENASE FROM STREPTOMYCES-AVERMITILIS AND THE PRODUCTION OF A FUNCTIONAL E1[ALPHA-BETA] COMPONENT IN ESCHERICHIA-COLI

被引：27

作者：

SKINNER, DD ^{[1
]}

MORGENSTERN, MR ^{[1
]}

FEDECHKO, RW ^{[1
]}

DENOYA, CD ^{[1
]}

机构：

[1] PFIZER INC, DIV CENT RES, BIOPROC RES, GROTON, CT 06340 USA

来源：

JOURNAL OF BACTERIOLOGY | 1995年 / 177卷 / 01期

关键词：

D O I：

10.1128/jb.177.1.183-190.1995

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A cluster of genes encoding the E1 alpha, E1 beta, and E2 subunits of branched chain alpha-keto acid dehydrogenase (BCDH) of Streptomyces avermitilis has been cloned and sequenced, Open reading frame 1 (ORF1) (E1 alpha), 1,146 nucleotides long, would encode a polypeptide of 40,969 Da (381 amino acids). ORF2 (E1 beta), 1,005 nucleotides long, would encode a polypeptide of 35,577 Da (334 amino acids). The intergenic distance between ORF1 and ORF2 is 73 bp. The putative ATG start codon of the incomplete ORF3 (E2) overlaps the stop codon of ORF2. Computer-aided searches showed that the deduced products of ORF1 and ORF2 resembled the corresponding E1 subunit (alpha or beta) of several prokaryotic and eukaryotic BCDH complexes. When these ORFs were overexpressed in Escherichia coli, proteins of about 41 and 34 kDa, which are the approximate masses of the predicted S. avermitilis ORF1 and ORF2 products, respectively, were detected. In addition, specific E1[alpha beta] BCDH activity was detected in E. coli cells carrying the S. avermitilis ORF1 (E1 alpha) and ORF2 (E1 beta) coexpressed under the control of the T7 promoter.

引用

页码：183 / 190

页数：8

共 72 条

[1] THE NUCLEOTIDE-SEQUENCE OF A 16S RIBOSOMAL-RNA GENE FROM STREPTOMYCES-COELICOLOR A3(2) [J].

BAYLIS, HA ;

BIBB, MJ .

NUCLEIC ACIDS RESEARCH, 1987, 15 (17) :7176-7176

[2] NUCLEOTIDE AND DEDUCED AMINO-ACID SEQUENCE OF THE ALPHA-SUBUNIT OF YEAST PYRUVATE-DEHYDROGENASE [J].

BEHAL, RH ;

BROWNING, KS ;

REED, LJ .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1989, 164 (02) :941-946

[3] THE M-GAMMA-DELTA-1 ELEMENT, A SMALL GAMMA-DELTA (TN1000) DERIVATIVE USEFUL FOR PLASMID MUTAGENESIS, ALLELE REPLACEMENT AND DNA SEQUENCING [J].

BERG, CM ;

VARTAK, NB ;

WANG, G ;

XU, XX ;

LIU, L ;

MACNEIL, DJ ;

GEWAIN, KM ;

WIATER, LA ;

BERG, DE .

GENE, 1992, 113 (01) :9-16

[4] GENE-EXPRESSION IN STREPTOMYCES - CONSTRUCTION AND APPLICATION OF PROMOTER-PROBE PLASMID VECTORS IN STREPTOMYCES-LIVIDANS [J].

BIBB, MJ ;

COHEN, SN .

MOLECULAR & GENERAL GENETICS, 1982, 187 (02) :265-277

[5] THE RELATIONSHIP BETWEEN BASE COMPOSITION AND CODON USAGE IN BACTERIAL GENES AND ITS USE FOR THE SIMPLE AND RELIABLE IDENTIFICATION OF PROTEIN-CODING SEQUENCES [J].

BIBB, MJ ;

FINDLAY, PR ;

JOHNSON, MW .

GENE, 1984, 30 (1-3) :157-166

[6] CLONING AND HETEROLOGOUS EXPRESSION IN STREPTOMYCES-LIVIDANS OF STREPTOMYCES-RIMOSUS GENES INVOLVED IN OXYTETRACYCLINE BIOSYNTHESIS [J].

BINNIE, C ;

WARREN, M ;

BUTLER, MJ .

JOURNAL OF BACTERIOLOGY, 1989, 171 (02) :887-895

[7]

BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513

[8]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[9] AVERMECTINS, NEW FAMILY OF POTENT ANTHELMINTIC AGENTS - PRODUCING ORGANISM AND FERMENTATION [J].

BURG, RW ;

MILLER, BM ;

BAKER, EE ;

BIRNBAUM, J ;

CURRIE, SA ;

HARTMAN, R ;

KONG, YL ;

MONAGHAN, RL ;

OLSON, G ;

PUTTER, I ;

TUNAC, JB ;

WALLICK, H ;

STAPLEY, EO ;

OIWA, R ;

OMURA, S .

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1979, 15 (03) :361-367

[10] SIMILARITY OF THE E1 SUBUNITS OF BRANCHED-CHAIN-OXOACID DEHYDROGENASE FROM PSEUDOMONAS-PUTIDA TO THE CORRESPONDING SUBUNITS OF MAMMALIAN BRANCHED-CHAIN-OXOACID AND PYRUVATE DEHYDROGENASES [J].

BURNS, G ;

BROWN, T ;

HATTER, K ;

IDRISS, JM ;

SOKATCH, JR .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 176 (02) :311-317

← 1 2 3 4 5 6 7 8 →