Cloning and expression of cDNA encoding mouse tyrosinase

Shigeki Shibahara, Yasushi Torruta, Teruyo Sakakura, Christoph Nager, Bhabatosh Chaudhuri, Rita Müller

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176 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2413-2427
Number of pages15
JournalNucleic acids research
Volume14
Issue number6
DOIs
Publication statusPublished - 1986 Mar 11

ASJC Scopus subject areas

  • Genetics

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    Shibahara, S., Torruta, Y., Sakakura, T., Nager, C., Chaudhuri, B., & Müller, R. (1986). Cloning and expression of cDNA encoding mouse tyrosinase. Nucleic acids research, 14(6), 2413-2427. https://doi.org/10.1093/nar/14.6.2413