摘要
主要通过建立Pygo2转基因小鼠的模型对其表型进行初步分析.首先构建K14-2×flag-Pygo的转基因构件,经酶切、纯化后构建Pygo2转基因小鼠.出生后的仔鼠用PCR和Western方法检测基因型,并通过进一步的免疫组化验证Pygo2基因的表达.PCR检测获得7只转基因阳性鼠,6只Western检测为阳性.对转基因小鼠子代的胚胎和成体进行免疫组化证明,Pygo2基因在皮肤和乳腺组织中有过量表达.转基因小鼠的皮肤、乳腺以及鼠尾椎骨等组织出现了异常的表型.乳腺中有肿瘤组织的形成,且Pygo2在肿瘤中有大量表达.该模型的成功建立为进一步研究Pygo2的功能奠定了基础.
Pygopus （Pygo） is a new component of Wnt signaling with conserved P HD domain, associates with Armadillo/β-catenin through the Legless/BCL9 adaptor. The objective of this study was to construct the specific expression of vector of Pygo2 and to establish a Pygo2 transgenie mouse model and cheek its phenotype. K14 promoter-2 × flag-Pygo2 construct was construeted,linearized, purified and then injected to produce transgenic mouse. The genotypes of transgenic founders were identified by PCR and Western. 71 offspring were born and 7 were proved to be positive founder. Protein expression of Pygo2 in tissues of transgenic mice were detected by immunohistochemistry. Pygo2 transgenic mice have been established successfully,6 in 7 viable offsprings were positive transgenic mice,the rate of positive mouse was 8. 45% （6/71）. In the skin and mammary gland of transgenic mice Pygo2 was over expressed. And there is abnormal phenotype in the skin,mammary gland and tail tissues in transgenie mice. Skin shows festering and abnormal phenotype of hair follicle,mammary gland shows early maturing and some transgenic mice occurs mammary gland tumor,including adenocarcinoma and adenoacanthoma. Pygo 2 protein is highly expressed in these tumor tissues. Taken together, these data indicate that the models will be contributed to the research of Pygo2 gene function.
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第2期266-271,共6页
Journal of Xiamen University：Natural Science
基金
国家自然科学基金(30671173)
福建省自然科学基金重点项目(2008J0007)
