Cloning and overexpression of a tyrosinase gene mel from Pseudomonas maltophila

Gelin Wang; Aberrahmane Aazaz; Zhenrong Peng; Ping Shen

doi:10.1016/S0378-1097(00)00065-3

Cloning and overexpression of a tyrosinase gene mel from Pseudomonas maltophila

Gelin Wang, Aberrahmane Aazaz, Zhenrong Peng, Ping Shen

Biochemistry

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

41 Scopus citations

Abstract

The tyrosinase gene (mel), which is responsible for melanin formation, was isolated by shotgun cloning of SalI fragments of Pseudomonas maltophila DNA. A 0.7-kb SalI fragment in the recombinant plasmid pWSY8 imparted the ability to synthesize melanin to an Escherichia coli host HB101. The nucleotide sequence of this DNA fragment revealed an open reading frame of 504 bp, encoding a protein of 169 amino acids. The fragment containing the mel gene was then cloned into an expression plasmid pPAS1 under the control of a promoter isolated from the host, P. maltophilia AT18. This strain increased the melanin production by 70.6% compared with the strain HB101/pWSY8, in which the cloned mel gene was under the control of the lac promoter from the vector pUC18. Copyright (C) 2000 Federation of European Microbiological Societies.

Original language	English (US)
Pages (from-to)	23-27
Number of pages	5
Journal	FEMS Microbiology Letters
Volume	185
Issue number	1
DOIs	https://doi.org/10.1016/S0378-1097(00)00065-3
State	Published - Apr 1 2000

Keywords

Melanin
Promoter
Pseudomonas maltophila
mel gene

ASJC Scopus subject areas

Microbiology
Molecular Biology
Genetics

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title = "Cloning and overexpression of a tyrosinase gene mel from Pseudomonas maltophila",

abstract = "The tyrosinase gene (mel), which is responsible for melanin formation, was isolated by shotgun cloning of SalI fragments of Pseudomonas maltophila DNA. A 0.7-kb SalI fragment in the recombinant plasmid pWSY8 imparted the ability to synthesize melanin to an Escherichia coli host HB101. The nucleotide sequence of this DNA fragment revealed an open reading frame of 504 bp, encoding a protein of 169 amino acids. The fragment containing the mel gene was then cloned into an expression plasmid pPAS1 under the control of a promoter isolated from the host, P. maltophilia AT18. This strain increased the melanin production by 70.6% compared with the strain HB101/pWSY8, in which the cloned mel gene was under the control of the lac promoter from the vector pUC18. Copyright (C) 2000 Federation of European Microbiological Societies.",

keywords = "Melanin, Promoter, Pseudomonas maltophila, mel gene",

author = "Gelin Wang and Aberrahmane Aazaz and Zhenrong Peng and Ping Shen",

note = "Funding Information: This work was supported by a grant 207960131 from the Science and Technology Commission of Hubei Province, PR China. We thank Lei Lei and Long Ma for their critical reading of the manuscript. ",

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AU - Wang, Gelin

AU - Aazaz, Aberrahmane

AU - Peng, Zhenrong

AU - Shen, Ping

N1 - Funding Information: This work was supported by a grant 207960131 from the Science and Technology Commission of Hubei Province, PR China. We thank Lei Lei and Long Ma for their critical reading of the manuscript.

PY - 2000/4/1

Y1 - 2000/4/1

N2 - The tyrosinase gene (mel), which is responsible for melanin formation, was isolated by shotgun cloning of SalI fragments of Pseudomonas maltophila DNA. A 0.7-kb SalI fragment in the recombinant plasmid pWSY8 imparted the ability to synthesize melanin to an Escherichia coli host HB101. The nucleotide sequence of this DNA fragment revealed an open reading frame of 504 bp, encoding a protein of 169 amino acids. The fragment containing the mel gene was then cloned into an expression plasmid pPAS1 under the control of a promoter isolated from the host, P. maltophilia AT18. This strain increased the melanin production by 70.6% compared with the strain HB101/pWSY8, in which the cloned mel gene was under the control of the lac promoter from the vector pUC18. Copyright (C) 2000 Federation of European Microbiological Societies.

AB - The tyrosinase gene (mel), which is responsible for melanin formation, was isolated by shotgun cloning of SalI fragments of Pseudomonas maltophila DNA. A 0.7-kb SalI fragment in the recombinant plasmid pWSY8 imparted the ability to synthesize melanin to an Escherichia coli host HB101. The nucleotide sequence of this DNA fragment revealed an open reading frame of 504 bp, encoding a protein of 169 amino acids. The fragment containing the mel gene was then cloned into an expression plasmid pPAS1 under the control of a promoter isolated from the host, P. maltophilia AT18. This strain increased the melanin production by 70.6% compared with the strain HB101/pWSY8, in which the cloned mel gene was under the control of the lac promoter from the vector pUC18. Copyright (C) 2000 Federation of European Microbiological Societies.

KW - Melanin

KW - Promoter

KW - Pseudomonas maltophila

KW - mel gene

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