The Tyrosinase Gene from Medakafish: Transgenic Expression Rescues Albino Mutation

Hidehito Inagaki, Akihiko Koga, Yoshitaka Bessho, Hiroshi Hori

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We have determined the 9.8 kb genomic nucleotide sequence of the tyrosinase gene and its 5′ upstream region from a teleost, medakafish (Oryzias latipes), and shown that the coding region is composed of five exons and four introns, spanning 4.7 kb. While the number and sizes of the exons were found to be similar to those of mammalian tyrosinase genes, however, the total size of the coding region (4.7 kb) was demonstrated to be less than one tenth those of mouse (ca. 70 kb) and human (> 70 kb) genes. Primer extension analysis revealed that the transcription initiation site starts with a long untranslated leader sequence (340 nucleotide long) from the AUG start codon. A characteristic CATGTG sequence known as a putative regulatory motif in melanocyte-specific genes was present in the 131st base upstream from the initiation site, while other typical regulatory elements such as the TATA-box or M-box common to terrestrial vertebrates were lacking. Transgenic experiments were carried out by microinjecting two kinds of plasmid clones into fertilized eggs of the albino i1 mutant: one consisting of the genomic tyrosinase gene with the 10 kb 5′ upstream region and the other the tyrosinase cDNA with the 3 kb 5′ upstream region. The results showed that 53 and 45 of 114 and 118 transgenic eggs, respectively, developed normally beyond hatching and 15 and 10 exhibited a mosaic pattern of pigmentation. Despite the absence of typical regulatory elements like a TATA-box in both cases correct melanin pigmentation was obtained without ectopic expression. Thus, transgenic expression rescued from the albino-i1 mutation, and the i locus of the medaka genome can be concluded to encode the tyrosinase gene.

Original languageEnglish
Pages (from-to)283-290
Number of pages8
JournalPigment Cell Research
Volume11
Issue number5
DOIs
Publication statusPublished - 01-01-1998

    Fingerprint

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Plant Science
  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this