Abstract:
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 pCS
2+ was digested with
BamHⅠ and
XhoⅠ restriction enzyme, after that
HOXC4 gene fragment was inserted into pCS
2+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-pCS
2+ 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.