Cloning of the histidine biosynthetic genes from Corynebacterium glutamicum: Organization and analysis of the hisG and hisE genes

2000 ◽  
Vol 46 (9) ◽  
pp. 848-855 ◽  
Author(s):  
Joon-Hye Kwon ◽  
Jae-Yeon Chun ◽  
Heung-Shick Lee ◽  
Choong-Ill Cheon ◽  
Eun-Sook Song ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Corynebacterium Glutamicum ◽  
Biosynthetic Pathway ◽  
Amino Acid Sequences ◽  
Sequencing Analysis ◽  
Gene Products ◽  
Gene Library ◽  
Biosynthetic Genes ◽  
Coding Regions ◽  
Genes Encoding

The physically linked hisG and hisE genes, encoding for ATP-phosphoribosyltransferase and phosphoribosyl-ATP-pyrophosphohydrolase were isolated from the Corynebacterium glutamicum gene library by complementation of Escherichia coli histidine auxotrophs. They are two of the nine genes that participate in the histidine biosynthetic pathway. Molecular genetics and sequencing analysis of the cloned 9-kb insert DNA showed that it carries the hisG and hisE genes. In combining this result with our previous report, we propose that all histidine biosynthetic genes are separated on the genome by three unlinked loci. The coding regions of the hisG and hisE genes are 279 and 87 amino acids in length with a predicted size of about 30 and 10 kDa, respectively. Computer analysis revealed that the amino acid sequences of the hisG and hisE gene products were similar to those of other bacteria.

Expression and Regulation of the Arsenic Resistance Operon of Acidiphilium multivorum AIU 301 Plasmid pKW301 in Escherichia coli

1998 ◽  
Vol 64 (2) ◽  
pp. 411-418 ◽  
Author(s):  
Katsuhisa Suzuki ◽  
Norio Wakao ◽  
Tetsuya Kimura ◽  
Kazuo Sakka ◽  
Kunio Ohmiya
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Constitutive Expression ◽  
Amino Acid Sequences ◽  
Gene Products ◽  
Arsenic Resistance ◽  
E Coli ◽  
Molecular Weights ◽  
Rna Polymerase Promoter ◽  
Extension Analysis

ABSTRACT The arsenic resistance (ars) operon from plasmid pKW301 of Acidiphilium multivorum AIU 301 was cloned and sequenced. This DNA sequence contains five genes in the following order: arsR, arsD, arsA,arsB, arsC. The predicted amino acid sequences of all of the gene products are homologous to the amino acid sequences of the ars gene products of Escherichia coliplasmid R773 and IncN plasmid R46. The ars operon cloned from A. multivorum conferred resistance to arsenate and arsenite on E. coli. Expression of the arsgenes with the bacteriophage T7 RNA polymerase-promoter system allowedE. coli to overexpress ArsD, ArsA, and ArsC but not ArsR or ArsB. The apparent molecular weights of ArsD, ArsA, and ArsC were 13,000, 64,000, and 16,000, respectively. A primer extension analysis showed that the ars mRNA started at a position 19 nucleotides upstream from the arsR ATG in E. coli. Although the arsR gene of A. multivorum AIU 301 encodes a polypeptide of 84 amino acids that is smaller and less homologous than any of the other ArsR proteins, inactivation of the arsR gene resulted in constitutive expression of the ars genes, suggesting that ArsR of pKW301 controls the expression of this operon.

scholarly journals Cloning and Sequence Analysis of Genes Encoding Staphylococcus hyicus Exfoliative Toxin Types A, B, C, and D

2004 ◽  
Vol 186 (6) ◽  
pp. 1833-1837 ◽  
Author(s):  
Peter Ahrens ◽  
Lars Ole Andresen
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Monoclonal Antibodies ◽  
Amino Acid ◽  
Sequence Analysis ◽  
Serine Proteases ◽  
Amino Acid Sequences ◽  
Coding Sequence ◽  
Exfoliative Toxin ◽  
Genes Encoding

ABSTRACT Exfoliative toxins produced by certain strains of Staphylococcus hyicus mediate exudative epidermitis in pigs. In this study the genes coding for four different exfoliative toxin from S. hyicus (ExhA, ExhB, ExhC, and ExhD) were cloned and sequenced. The coding sequence of the four toxin genes ranged from 816 to 834 bp. The amino acid sequences of these four toxins were homologous to the earlier described exfoliative toxins SHETB from S. hyicus and ETA, ETB, and ETD from Staphylococcus aureus. The homology between the S. hyicus toxins was at the same level as the homology to the exfoliative toxins from S. aureus. The toxins showed similarity to serine proteases, including preservation of the catalytic tract in ExhA, ExhB, and ExhC. However, in ExhD, Asp in the putative catalytic tract was replaced with Glu. The recombinant toxins could be expressed in Escherichia coli, and three of the four toxins were recognized by monoclonal antibodies raised against native exfoliative toxins.

Cloning and analysis of the aroB gene encoding dehydroquinate synthase from Corynebacterium glutamicum

1999 ◽  
Vol 45 (10) ◽  
pp. 885-890 ◽  
Author(s):  
Min-Ah Han ◽  
Heung-Shick Lee ◽  
Choong-Ill Cheon ◽  
Kyung-Hee Min ◽  
Myeong-Sok Lee
Keyword(s):  
Escherichia Coli ◽  
Corynebacterium Glutamicum ◽  
Auxotrophic Mutant ◽  
Gene Products ◽  
Gene Encoding ◽  
Reading Frame ◽  
E Coli ◽  
Dna Library ◽  
Genomic Dna Library ◽  
Dehydroquinate Synthase

The aroB gene encoding dehydroquinate synthase of Corynebacterium glutamicum has been cloned by complementation of an aro auxotrophic mutant of Escherichia coli with the genomic DNA library. The recombinant plasmid contained a 1.4-kb fragment that complemented the Escherichia coli dehydroquinate-synthase-deficient mutant. The nucleotide sequences of the subcloned DNA has been determined. The sequences contain an open reading frame of 360 codons, from which a protein with a molecular mass of about 38 kDa could be predicted. This is consistent with the size of the AroB protein expressed in E. coli. Alignment of different prokaryotic and eukaryotic aroB gene products reveals an overall identity ranging from 29 to 57% and the presence of several highly conserved regions.Key words: Corynebacterium glutamicum, aromatic amino acid biosynthetic gene, dehydroquinate synthase, aroB gene.

scholarly journals Colistin-Resistant mcr-1-Positive Escherichia coli ST131-H22 Carrying blaCTX–M–15 and qnrB19 in Agricultural Soil

2021 ◽  
Vol 12 ◽  
Author(s):  
Ralf Lopes ◽  
João Pedro Rueda Furlan ◽  
Lucas David Rodrigues dos Santos ◽  
Inara Fernanda Lage Gallo ◽  
Eliana Guedes Stehling
Keyword(s):  
Escherichia Coli ◽  
Agricultural Soil ◽  
Meat Products ◽  
Environmental Safety ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Sequencing Analysis ◽  
E Coli ◽  
Clade B ◽  
Genes Encoding

The pandemic Escherichia coli sequence type 131 (ST131) carrying plasmid-mediated colistin resistance mcr genes has emerged worldwide causing extraintestinal infections, with lineages belonging to three major clades (A, B, and C). Clade B is the most prevalent in animals, contaminating associated meat products, and can be transmitted zoonotically. However, the blaCTX–M–15 gene has only been associated with C2 subclade so far. In this study, we performed a genomic investigation of an E. coli (strain S802) isolated from a kale crop in Brazil, which exhibited a multidrug-resistant (MDR) profile to clinically significant antimicrobials (i.e., polymyxin, broad-spectrum cephalosporins, aminoglycosides, and fluoroquinolones). Whole-genome sequencing analysis revealed that the S802 strain belonged to serotype O25:H4, ST131/CC131, phylogenetic group B2, and virotype D5. Furthermore, S802 carried the clade B-associated fimH22 allele, genes encoding resistance to clinically important antimicrobials, metals, and biocides, and was phylogenetically related to human, avian, and swine ST131-H22 strains. Additionally, IncHI2-IncQ1, IncF [F2:A-:B1], and ColE1-like plasmids were identified harboring mcr-1.1, blaCTX–M–15, and qnrB19, respectively. The emergence of the E. coli ST131-H22 sublineage carrying mcr-1.1, blaCTX–M–15, and qnrB19 in agricultural soil represents a threat to food and environmental safety. Therefore, a One Health approach to genomic surveillance studies is required to effectively detect and limit the spread of antimicrobial-resistant bacteria and their resistance genes.

scholarly journals Cloning and Expression of clt Genes Encoding Milk-Clotting Proteases from Myxococcus xanthus 422

2004 ◽  
Vol 70 (10) ◽  
pp. 6337-6341 ◽  
Author(s):  
M. Poza ◽  
M. Prieto-Alcedo ◽  
C. Sieiro ◽  
T. G. Villa
Keyword(s):  
Escherichia Coli ◽  
Myxococcus Xanthus ◽  
Open Reading Frames ◽  
Cloning And Expression ◽  
Gene Library ◽  
Expression Systems ◽  
E Coli ◽  
Milk Clotting ◽  
Genes Encoding ◽  
Reading Frames

ABSTRACT The screening of a gene library of the milk-clotting strain Myxococcus xanthus 422 constructed in Escherichia coli allowed the description of eight positive clones containing 26 open reading frames. Only three of them (cltA, cltB, and cltC) encoded proteins that exhibited intracellular milk-clotting ability in E. coli, Saccharomyces cerevisiae, and Pichia pastoris expression systems.

scholarly journals Identification and Expression of the Genes Encoding a Reactivating Factor for Adenosylcobalamin-Dependent Glycerol Dehydratase

1999 ◽  
Vol 181 (13) ◽  
pp. 4110-4113 ◽  
Author(s):  
Takamasa Tobimatsu ◽  
Hideki Kajiura ◽  
Michio Yunoki ◽  
Muneaki Azuma ◽  
Tetsuo Toraya
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Open Reading Frames ◽  
Gene Products ◽  
Glycerol Dehydratase ◽  
Permeabilized Cells ◽  
Genes Encoding ◽  
Reading Frames

ABSTRACT Adenosylcobalamin-dependent glycerol dehydratase undergoes inactivation by glycerol, the physiological substrate, during catalysis. In permeabilized cells of Klebsiella pneumoniae, the inactivated enzyme is reactivated in the presence of ATP, Mg2+, and adenosylcobalamin. We identified the two open reading frames as the genes for a reactivating factor for glycerol dehydratase and designated them gdrA and gdrB. The reactivation of the inactivated glycerol dehydratase by the gene products was confirmed in permeabilized recombinant Escherichia coli cells coexpressing GdrA and GdrB proteins with glycerol dehydratase.

scholarly journals Genome Mining Reveals Two Missing CrtP and AldH Enzymes in the C30 Carotenoid Biosynthesis Pathway in Planococcus faecalis AJ003T

Molecules ◽  
2020 ◽  
Vol 25 (24) ◽  
pp. 5892
Author(s):  
Jun Ho Lee ◽  
Jin Won Kim ◽  
Pyung Cheon Lee
Keyword(s):  
Escherichia Coli ◽  
Biosynthetic Pathway ◽  
Genome Mining ◽  
Carotenoid Biosynthesis ◽  
Biosynthesis Pathway ◽  
Gene Encoding ◽  
A Genome ◽  
Genes Encoding ◽  
Carotenoid Biosynthesis Pathway ◽  
Carotenoid Pathway

Planococcus faecalis AJ003T produces glycosyl-4,4′-diaponeurosporen-4′-ol-4-oic acid as its main carotenoid. Five carotenoid pathway genes were presumed to be present in the genome of P. faecalis AJ003T; however, 4,4-diaponeurosporene oxidase (CrtP) was non-functional, and a gene encoding aldehyde dehydrogenase (AldH) was not identified. In the present study, a genome mining approach identified two missing enzymes, CrtP2 and AldH2454, in the glycosyl-4,4′-diaponeurosporen-4′-ol-4-oic acid biosynthetic pathway. Moreover, CrtP2 and AldH enzymes were functional in heterologous Escherichia coli and generated two carotenoid aldehydes (4,4′-diapolycopene-dial and 4,4′-diaponeurosporene-4-al) and two carotenoid carboxylic acids (4,4′-diaponeurosporenoic acid and 4,4′-diapolycopenoic acid). Furthermore, the genes encoding CrtP2 and AldH2454 were located at a distance the carotenoid gene cluster of P. faecalis.

scholarly journals Genome-wide transcriptome analysis of triterpene biosynthetic genes of Anoectochilus roxburghii plant

2020 ◽  
Author(s):  
Hongzhen Wang ◽  
Haishun Xu ◽  
Peter E. Brodelius ◽  
Xueqian Wu ◽  
qingsong Shao ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Sequence Similarity ◽  
Bioactive Components ◽  
Biosynthetic Genes ◽  
Bioactive Component ◽  
Genome Wide ◽  
Three Samples ◽  
Anoectochilus Roxburghii ◽  
Genes Encoding ◽  
Cellular Components

Abstract Background: Anoectochilus roxburghii is a medicinal plant and contains a variety of bioactive components, including triterpene, which exhibits important pharmacological properties with low toxicity. However, little is known about the biosynthetic pathway of triterpene or about the genome and transcriptome in A. roxburghii. Results: In order to analyze transcriptional determinants related to the biosynthesis of the bioactive components, we performed transcriptome sequencing in A. roxburghii (SRX1818644, SRX1818642 and SRX1818641) and annotated the sequences from three samples. In total, 137,679,059 clean reads were obtained, corresponding to 12.20 Gb of total nucleotides. They were then assembled into 86,382 contigs and 68,938 unigenes, which were further annotated according to sequence similarity with known genes in COG, EST, Nr, Pfam and Uniprot databases, leading to 10,040，29,442，39,551，34,991 and 28,082 unigenes, respectively. GO analysis classified all unigenes into three functional categories, i.e. biological processes (43,206 unigenes in 22 categories), molecular functions (46,978 unigenes in 15 categories) and cellular components (20,951 unigenes in 18 categories). Candidate triterpenes biosynthetic genes ArHMGR1 in MEV pathway, ArDXS1, ArDXS4 ArDXS5, ArDXS8-10, ArDXR1-2 and ArHDR1-2 in MEP pathway and ArFDS1, ArSM and ArOCS were selected based on RNA-seq and gene-to-metabolites correlation analysis. Conclusion: The transcriptomes of A. roxburghii plant include 86,382 contigs and 68,938 unigenes. The assembled dataset allowed identification of genes encoding enzymes in the biosynthesis of bioactive components in A. roxburghii plant. Candidate genes that encode enzymes being important in triterpenes biosynthetic pathway were selected. This will facilitate the study of expression and regulation in the biosynthesis of bioactive component in A.roxburghii.

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