Nucleic Acids Research, 1986, Vol. 14, No. 6 2413-2427
© 1986
Articles |
Cloning and expression of cDNA encoding mouse tyrosinase
,Friedrich Miescher-Institut PO Box 2543, CH-002 Basel, Switzerland 1Department of Dermatology, Tohoku University School of Medicine Sendai, Miyagi 980, Japan
*Aichi Cancer Center Research Institute, Nagoya, Japan
+Ciba-Geigy Ltd., Basel, Switzerland
To whom reprint requests should be sent.
Received January 30, 1986. Accepted February 26, 1986.
We have isolated a pigment cell-specific cDNA clone from a B16 mouse melanoma cDNA library by differential hybridization. The mRNA of isolated cDNA is highly expressed in B16 melanoma cells and in black mouse (C57BL/6) skin, but is not detectable in mouse neuroblastoma cells nor in K1735 mouse amelanotic melanoma cells. The protein sequence deduced from the nucleotide sequence of the cloned cDNA shows significant similarity to the entire region of Neurospora tyrosinase. To know the identity of cDNA, we transfected K1735 amelanotic melanoma and COS-7 cells with the cDNA carried in a simian virus 40 vector (pKCRH2). We confirmed that the isolated cDNA encodes mouse tyrosinase by immunofluorescence staining of transfected cells using two different anti-T4-tyrosinase monoclonal antibodies. Tyrosinase is composed of 513 amino acids with a molecular weight of 57,872 excluding a hydrophobic signal peptide of 24 amino acids.
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