A cDNA clone for human tyrosinase

Details A cDNA clone for human tyrosinase A cDNA clone for human tyrosinase

1986-10-06

1990-02-06

Mays, Thomas D.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435189, 4351723, 435235, 435320, 536 27, 935 14, 935 31, 935 78, C12Q 168, C12N 902, C12N 1500, C07H 1512

Patent

active

048988146

ABSTRACT:
This discovery resulted when a .lambda. gt11 cDNA library of normal human melanocytes were screened with antibodies directed against purified hamster tyrosinase. Sixteen independent clones which gave a positive signal were isolated from 5.times.10.sup.5 independent plaques. cDNA inserts of 13 clones among the 16 candidates cross-hybridized with each other, indicating that they were from related mRNA species. mRNA homologous to a representative cDNA .lambda. mel 34 was expressed specifically in melanocytes, detecting an approximately 2.4 kb mRNA species of human melanocytes. The nucleotide sequence of the three overlapping cDNA inserts spanning 1.88 kb was determined and an amino acid sequence was deducted. The human tyrosinase is composed of 548 amino acids with a molecular weight of 62,160 excluding a hydrophobic signal peptide. Mouse genomic DNA blot analysis revealed that the gene for .lambda. mel 34 was deleted in albino mouse homozygous for the deletion at and around the albino locus on chromosome 7. It is concluded that .lambda. mel 34 contained cDNA encoding human tyrosinase. Moreover, the cDNA gene for human tyrosinase was recovered from the .lambda. mel 34 and can be used for many purposes including the production of pure human tyrosinase.

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