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The zebrafish erythropoietin: functional identification and biochemical characterization
Authors:Chu Cheng-Ying  Cheng Chia-Hsiung  Chen Gen-Der  Chen Yi-Chung  Hung Chin-Chun  Huang Kai-Yun  Huang Chang-Jen
Affiliation:Graduate Institute of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan.
Abstract: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 alphaA1- and betaA1-globin, but not the embryonic betae1-globin, was affected.
Keywords:Erythropoietin   Zebrafish   Morpholino oligonucleotide   Glycoprotein   Secretion   Globin staining
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