Expression of HOXC4 Gene during Early Development in Zebrafish

Objecitve To investigate the mechanism of HOXC4 gene during early development in zebrafish, study it's expression in hematopoiesis system with whole mount in situ hybridization(WISH), then make a foundation work for further study of HOXC4 gene in hematopoiesis development. Methods The total RNA of different phases of zebrafish embryos was extracted, and a fragment of HOXC4 gene was cloned with RT-PCR, then HOXC4 gene fragment and pCS²⁺ was digested with BamHⅠ and XhoⅠ restriction enzyme, after that HOXC4 gene fragment was inserted into pCS²⁺vector. After confirming the constructed plasmid, the digoxingenin-labeled anti-sence mRNA probe of HOXC4 gene was synthesized in vitro by using T3 RNA polymerase, the exprression pattern of HOXC4 during zebrafish embryogenesis using anti-sence probe with WISH was conducted. Results HOXC4 gene was cloned by RT-PCR and HOXC4-pCS²⁺ plasmid was constructed and confirmed, expression of HOXC4 showed that it was expressed highly in nervous system of wild-type (WT) Tuebingen zebrafish. Conculsion The expression pattern of HOXC4 during early development in zebrafish was obtained, and it is very important for further study of HOXC4 gene function in zebrafish.