Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium

Ashok Kumar Chopra; Johnny Wayne Peterson; Rajendra Prasad

doi:10.1016/0167-4781(91)90224-A

Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium

Ashok Kumar Chopra, Johnny Wayne Peterson, Rajendra Prasad

Microbiology And Immunology

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

The nucleotide sequence of the hydHG operon, comprised of chromosomal genes that regulate labile hydrogenase activity in Salmonella typhimurium, was compared with the reported hydHG sequence of Escherichia coli. Nucleotide sequence analysis of a 4.8 kb EcoRI fragment of Salmonella chromosomal DNA revealed that one of the open reading frames (ORF) encoded a protein of 441 amino acid residues. This large ORF was identified on a 2.7 kb EcoRI/HindIII fragment and coded for the complete hydG gene. The carboxy-terminus (626 bp) of the hydH gene also was present immediately upstream of hydG. Expression of the Salmonella hydG gene in a T7 promoter/ polymerase system revealed the presence of a unique 45 kDa protein band. The incomplete hydH gene was not expressed. It is proposed that the labile hydrogenase activity in S. typhimurium may be regulated by the multiple component system.

Original language	English (US)
Pages (from-to)	115-118
Number of pages	4
Journal	BBA - Gene Structure and Expression
Volume	1129
Issue number	1
DOIs	https://doi.org/10.1016/0167-4781(91)90224-A
State	Published - Dec 2 1991

Keywords

DNA sequencing
T7 polymerase/promoter system
hydG gene
hydH gene

ASJC Scopus subject areas

Structural Biology
Biophysics
Biochemistry
Genetics

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title = "Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium",

abstract = "The nucleotide sequence of the hydHG operon, comprised of chromosomal genes that regulate labile hydrogenase activity in Salmonella typhimurium, was compared with the reported hydHG sequence of Escherichia coli. Nucleotide sequence analysis of a 4.8 kb EcoRI fragment of Salmonella chromosomal DNA revealed that one of the open reading frames (ORF) encoded a protein of 441 amino acid residues. This large ORF was identified on a 2.7 kb EcoRI/HindIII fragment and coded for the complete hydG gene. The carboxy-terminus (626 bp) of the hydH gene also was present immediately upstream of hydG. Expression of the Salmonella hydG gene in a T7 promoter/ polymerase system revealed the presence of a unique 45 kDa protein band. The incomplete hydH gene was not expressed. It is proposed that the labile hydrogenase activity in S. typhimurium may be regulated by the multiple component system.",

keywords = "DNA sequencing, T7 polymerase/promoter system, hydG gene, hydH gene",

author = "Chopra, {Ashok Kumar} and Peterson, {Johnny Wayne} and Rajendra Prasad",

note = "Funding Information: This work was supported by NIH grant AI 18401-09 from NIAID. The authors acknowledge the gift of E. coli cultures containing plasmids pGPI-2, pT7-5 and pT7-6 received from Dr. S. Tabor, Harvard Medical School, Boston, MA, Excellent technical assistance of Tuyet Vo during sequencing of the hydHG genes is greatly appreciated.",

year = "1991",

month = dec,

day = "2",

doi = "10.1016/0167-4781(91)90224-A",

language = "English (US)",

volume = "1129",

pages = "115--118",

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T1 - Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium

AU - Chopra, Ashok Kumar

AU - Peterson, Johnny Wayne

AU - Prasad, Rajendra

N1 - Funding Information: This work was supported by NIH grant AI 18401-09 from NIAID. The authors acknowledge the gift of E. coli cultures containing plasmids pGPI-2, pT7-5 and pT7-6 received from Dr. S. Tabor, Harvard Medical School, Boston, MA, Excellent technical assistance of Tuyet Vo during sequencing of the hydHG genes is greatly appreciated.

PY - 1991/12/2

Y1 - 1991/12/2

N2 - The nucleotide sequence of the hydHG operon, comprised of chromosomal genes that regulate labile hydrogenase activity in Salmonella typhimurium, was compared with the reported hydHG sequence of Escherichia coli. Nucleotide sequence analysis of a 4.8 kb EcoRI fragment of Salmonella chromosomal DNA revealed that one of the open reading frames (ORF) encoded a protein of 441 amino acid residues. This large ORF was identified on a 2.7 kb EcoRI/HindIII fragment and coded for the complete hydG gene. The carboxy-terminus (626 bp) of the hydH gene also was present immediately upstream of hydG. Expression of the Salmonella hydG gene in a T7 promoter/ polymerase system revealed the presence of a unique 45 kDa protein band. The incomplete hydH gene was not expressed. It is proposed that the labile hydrogenase activity in S. typhimurium may be regulated by the multiple component system.

AB - The nucleotide sequence of the hydHG operon, comprised of chromosomal genes that regulate labile hydrogenase activity in Salmonella typhimurium, was compared with the reported hydHG sequence of Escherichia coli. Nucleotide sequence analysis of a 4.8 kb EcoRI fragment of Salmonella chromosomal DNA revealed that one of the open reading frames (ORF) encoded a protein of 441 amino acid residues. This large ORF was identified on a 2.7 kb EcoRI/HindIII fragment and coded for the complete hydG gene. The carboxy-terminus (626 bp) of the hydH gene also was present immediately upstream of hydG. Expression of the Salmonella hydG gene in a T7 promoter/ polymerase system revealed the presence of a unique 45 kDa protein band. The incomplete hydH gene was not expressed. It is proposed that the labile hydrogenase activity in S. typhimurium may be regulated by the multiple component system.

KW - DNA sequencing

KW - T7 polymerase/promoter system

KW - hydG gene

KW - hydH gene

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JO - Biochimica et Biophysica Acta - Gene Structure and Expression

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ER -

Cloning and sequence analysis of hydrogenase regulatory genes (hydHG) from Salmonella typhimurium

Abstract

Keywords

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