Molecular cloning and functional expression of feline thrombopoietin

Haruka Matsushiro, Hirotomo Kato, Tomoyuki Tahara, Takashi Kato, Akira Iwata, Toshihiro Watari, Hajime Tsujimoto, Atsuhiko Hasegawa

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Feline thrombopoietin (TPO) was molecularly cloned to establish a basis for cytokine therapy of thrombocytopenia in cats. cDNA clones covering the whole coding sequence of feline TPO were isolated from feline liver. The feline TPO cDNA obtained in this study contained an open reading frame encoding 349 amino acid residues. The predicted amino acid sequence of feline TPO shared 78.7, 69.9, 72.9 and 83.0% similarity with sequences of human, murine, rat and canine TPO, respectively. Four cysteine residues and two of four N-glycosylation sites that are conserved among species were also found at the corresponding positions in feline TPO. The feline TPO cDNA fragment encoding the whole amino acid coding region was recloned into an expression vector, and the resulting vector was transfected into 293T cells using the calcium phosphate method. The supernatant of the transfected 293T cells stimulated the proliferation of a human megakaryoblastic leukemia cell line (UT-7/TPO) cells in a dose dependent manner, indicating that the feline TPO cDNA obtained in this study encodes biologically active feline TPO.

Original languageEnglish
Pages (from-to)225-236
Number of pages12
JournalVeterinary Immunology and Immunopathology
Volume66
Issue number3-4
DOIs
Publication statusPublished - 1998 Dec 11

Keywords

  • Feline
  • Molecular cloning
  • Thrombopoietin

ASJC Scopus subject areas

  • Immunology
  • veterinary(all)

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  • Cite this

    Matsushiro, H., Kato, H., Tahara, T., Kato, T., Iwata, A., Watari, T., Tsujimoto, H., & Hasegawa, A. (1998). Molecular cloning and functional expression of feline thrombopoietin. Veterinary Immunology and Immunopathology, 66(3-4), 225-236. https://doi.org/10.1016/S0165-2427(98)00190-1