斑马鱼Shh、Wnt5b和Catnb特异性基因片段的克隆

Molecular cloning of Shh,Wnt5b and Catnb Specific Gene Fragments from Zebrafish Embryos

目的:克隆斑马鱼Shh(sonic hedgehox)、Wnt5b(wingless-related MMTV integration site 5b)和Catnb(β-catenin)特异性基因片段。方法:选取发育3d的斑马鱼胚胎,利用RT-PCR技术直接克隆Shh、Wnt5b和Catnb基因特异性基因片段,将其亚克隆入pGM-T载体进行序列测定。结果:从发育3d的斑马鱼胚胎中分别获得511bp、551bp和424bp的特异性片段。结论:序列分析表明,其与Gen-Bank中已公布的斑马鱼Shh、Wnt5b和Catnb基因序列一致。从发育3d的斑马鱼胚胎中成功地克隆了斑马鱼Shh、Wnt5b和Catnb特异性基因序列片段。

Objective：To clone the specific gene fragments of Shh,Wnt5b and Catnb genes from zebrafish embryos. Methods：The third day postnatal embryos of zebrafish were collected and after their heads were sectioned the total RNA were extracted. The specific gene fragments of Shh,Wnt5b and Catnb from zebrafish embryos were cloned by RT-PCR. The target fragments were inserted into the pGM-T vector and sequenced. Results：The 511bp,551bp and 424bp fragments were obtained from zebrafish embryos. The sequence analysis showed that they were 99.9% homologous to zabrafish Shh,Wnt5b and Catnb specific gene reported by Gen-Bank respectively. Conclusion：The specific fragments of Shh,Wnt5b and Catnb of zebrafish were successfully cloned from zebrafish embryos.