CAJ | 학술논문

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.
주요통과건립Pygo2전기인소서적모형대기표형진행초보분석.수선구건K14-2×flag-Pygo적전기인구건,경매절、순화후구건Pygo2전기인소서.출생후적자서용PCR화Western방법검측기인형,병통과진일보적면역조화험증Pygo2기인적표체.PCR검측획득7지전기인양성서,6지Western검측위양성.대전기인소서자대적배태화성체진행면역조화증명,Pygo2기인재피부화유선조직중유과량표체.전기인소서적피부、유선이급서미추골등조직출현료이상적표형.유선중유종류조직적형성,차Pygo2재종류중유대량표체.해모형적성공건립위진일보연구Pygo2적공능전정료기출.
Pygopus (Pygo) is a new component of Wnt signaling with conserved PHD domain,associates with Armadillo/ nstructed,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 transgenic 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.