RESUMO
OBJECTIVE: 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 BamH I and Xho I 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. CONCLUSION: 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.
