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The zebrafish erythropoietin: Functional identification and biochemical characterization
Author(s) -
Chu Cheng-Ying,
Cheng Chia-Hsiung,
Chen Gen-Der,
Chen Yi-Chung,
Hung Chin-Chun,
Huang Kai-Yun,
Huang Chang-Jen
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.07.073
Subject(s) - zebrafish , erythropoietin , complementary dna , globin , microbiology and biotechnology , morpholino , biology , transfection , grass carp , recombinant dna , messenger rna , cell culture , gene , cloning (programming) , biochemistry , genetics , fishery , fish <actinopterygii> , computer science , programming language
In the present study, the zebrafish epo cDNA was cloned. The encoded protein displays 90%, 55% and 32% identity to the Epo from carp, fugu and human, respectively. Through RT‐PCR, the expression of zepo mRNA was mainly in the heart and liver. In the COS‐1 cell transfection experiments, the recombinant zEpo‐HA protein was efficiently secreted into the culture medium as a glycoprotein and the carbohydrate moiety can be cleaved by the treatment of peptide‐ N ‐glycosidase F (PNGase F). Using the morpholino approach, we showed that zepo morphants displayed severe anemia leading to high mortality during development. Such an effect can be significantly rescued by zepo RNA. Furthermore, in the absence of functional zEpo, the expression of specific markers for adult globin genes, such as α A1 ‐ and β A1‐globin , but not the embryonic β e1‐globin , was affected.