肝脏特异HPPCn转基因小鼠的建立及表型分析  被引量:1

Establishment and phenotypic analysis of liver-specific over-expression HPPCn transgenic mice

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作  者:文川[1] 赛岩[2] 白纪民[2] 刘勇[2] 常菁[2] 王智[1] 崔春萍[2] 

机构地区:[1]湖南农业大学生物科学技术学院,长沙410128 [2]军事医学科学院放射与辐射研究所,北京100850

出  处:《中国医药生物技术》2012年第2期100-105,共6页Chinese Medicinal Biotechnology

基  金:国家自然科学基金(30930041)

摘  要:目的建立高表达HPPCn转基因小鼠,为研究该基因在肝癌发生发展中的功能提供研究模型。方法显微注射外源基因pGEM-A1b/his-HPPCn至FVB/N小鼠原核,注射受精卵移植到假孕受体出生个体。PCR和Southern blot鉴定转基因小鼠基因型,Western blot和Realtime-PCR检测转基因阳性和阴性小鼠肝脏HPPCn蛋白表达情况,HE染色和透射电镜观察肝脏病理改变。结果经检测,显微注射共产生2只首建鼠,转入的重组HPPCn基因能在肝脏中特异表达,并能够稳定遗传。转基因小鼠肝脏内HPPCn含量明显增高。转基因后并未对小鼠造成不良影响。结论 HPPCn参与了肝癌发生发展的过程,其在体内的高水平表达,为在体内研究该基因的功能提供了工具。Objective To generate the high level expression transgenic HPPCn mice, an animal model for the study of its function and effects on Hepatocellular carcinoma. Methods Transgenic mice were generated by pro-nuclear microinjection and embryo transplantation. Positive founder mice were identified by using both PCR and Southern blot. The expression of HPPCn in the liver tissues from wild type mice or transgenic mice was analyzed by using Western blot and realtime-PCR. The pathologic changes of the liver were analyzed by TEM and microscopy. Results Two fotmder transgenic mice were obtained by microinjection, exogenous HPPCn gene existed in the liver tissues, and stable passage of the gene in the transgenic mice was achieved. The expression of HPPCn was up-regulated in the liver tissue of transgenic mice and there was no adverse effect on mice. Conclusions HPPCn is involved in the occurrence and development of hepatocellular carcinoma. When its expression is raised, it can be used to investigate the effect of HPPCn gene in vivo levels.

关 键 词:小鼠 转基因 肝肿瘤 白细胞介素6 

分 类 号:R-332[医药卫生]

 

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