Cloning and expression of tyrosinase gene from Pseudomonas maltophilia in E. coli

Ge Lin Wang, Ping Shen, Lan Yang, Zhen Rong Peng

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

The enzyme tyrosinase, encoded by tyrosinase gene (mel), is responsible for melanin formation. In a shotgun cloning experiment, a SalI-digested DNA fragment coding for tyrosinase was cloned from Pseudomonas maltophilia DNA into plasmid vector (pUC18) to generate the hybrid plasmid (pWSY). The recombinant plasmid imparted the ability of melanin synthesis to an E. coli host (HB010). The foreign DNA fragment (0.7kb) possessed no recognition sites for BamHI, HindIII, EcoRI or BclI. Hybridization studies confirmed that the small fragment cloned in pWSY was from P. maltophilia DNA. Nucleotide sequence analysis identified an ORF of 504nt coding tyrosinase. SDS-PAGE analysis also revealed an additional protein of 18kDa, which was equal to the putative tyrosinase according to the size of mel fragment, was expressed in the E. coli recombinant carrying the plasmid pWSY.

Original languageEnglish (US)
Pages (from-to)278-279
Number of pages2
JournalActa Genetica Sinica
Volume26
Issue number3
StatePublished - Dec 1 1999

Keywords

  • Cloning
  • Expression
  • Mel
  • Pseudomonas maltophilia
  • Sequencing

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Wang, G. L., Shen, P., Yang, L., & Peng, Z. R. (1999). Cloning and expression of tyrosinase gene from Pseudomonas maltophilia in E. coli. Acta Genetica Sinica, 26(3), 278-279.