Open AccessCloning and overexpression of a tyrosinase gene mel from Pseudomonas maltophila
Author(s) -
Wang Gelin,
Aazaz Aberrahmane,
Peng Zhenrong,
Shen Ping
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09035.x
Subject(s) - microbiology and biotechnology , plasmid , molecular cloning , biology , open reading frame , gene , cloning (programming) , nucleic acid sequence , tyrosinase , pseudomonas , escherichia coli , recombinant dna , cloning vector , melanin , peptide sequence , genetics , biochemistry , bacteria , enzyme , computer science , programming language
The tyrosinase gene ( mel ), which is responsible for melanin formation, was isolated by shotgun cloning of Sal I fragments of Pseudomonas maltophila DNA. A 0.7‐kb Sal I 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.