斑马鱼甲状腺球蛋白基因的克隆及其组织表达

目的克隆斑马鱼甲状腺球蛋白（TG）基因，并研究TG在斑马鱼组织的表达状况。方法克隆TG基因，制备DIG标记的RNA探针，采用全胚胎原位杂交方法研究TG基因在胚胎斑马鱼的表达；构建融合蛋白表达载体pGEX-TG，制备多克隆抗体，应用免疫组化方法研究TG在成年斑马鱼甲状腺组织的表达。结果成功克隆了斑马鱼TG基因（GenBank登录号Dq278875），利用全胚胎原位杂交及免疫组化方法证实TG基因在胚胎期及成年期斑马鱼甲状腺组织均特异表达。结论TG特异性地在胚胎期和成年期的斑马鱼甲状腺组织表达，斑马鱼是研究甲状腺发育调控分子机制的有优势的模式生物。

Cloning and expression of thyroglobulin gene in thyroid gland of zebrafish

ObjectiveTo clone the thyroglobulin (TG) gene of zebrafish and investigate TG gene expression in zebrafish during embryonic development and in adult. MethodsTG cDNA of zebrafish was cloned and its digoxin-labeled RNA probe was prepared. The expression pattern of TG gene in embryonic zebrafish was investigated by whole embryo mount in situ hybridization. Furthermore, TG protein of zebrafish was expressed in the bacteria via pGET-TG construction and the polyclonal antibody for TG protein was prepared. The expression of TG protein in thyroid gland of adult zebrafish was studied by immunohistochemistry. ResultsTG gene of zebrafish was successfully cloned(GenBank Accession Number:Dq278875). By in whole embryo mount in situ hybridization and immunohistochemistry,it was found that TG was specifically expressed in thyroid gland of zebrafish during embryonic development and in adult. ConclusionTG gene is specifically expressed in thyroid gland of zebrafish during embryonic development and in adult. The conservation of the TG gene expression and protein structure from zebrafish to human supports the view that zebrafish is a favourite model organism for studying thyroid development in vertebrates.