BAT3过表达提高内源朊蛋白的表达量  

Overexpression of BAT3 enhanced cellular levels of endogenous prion protein

作　　者：宋志琦[1] 杨利峰[1] 王运盛[1] 朱婷[1] 赵德明[1] 

机构地区：[1]中国农业大学动物医学院国家动物海绵状脑病实验室,北京海淀100193

出　　处：《中国兽医杂志》2015年第11期9-11,15,I0001,共5页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金(31272532)

摘　　要：为了检测BAT3蛋白与朊蛋白的相互作用,探讨BAT3蛋白在朊蛋白合成和降解以及海绵状脑病发生过程中所起的作用。构建能够在细胞中表达全长朊蛋白的真核表达载体pGEFP-N1-PRNP和表达全长BAT3蛋白的真核表达载体pcDNA3.1-HA-BAT3,应用脂质体方法分别将质粒转染进入Hela细胞,原代神经元,Neuro2a细胞。利用免疫荧光技术观察内源BAT3与全长朊蛋白的相互作用情况,利用免疫印迹技术检测BAT3过表达对内源朊蛋白表达的影响。结果在Hela细胞和原代神经元内,转染pGEFP-N1-PRNP后,自发绿色荧光蛋白的全长朊蛋白与内源BAT3发生共定位;Hela细胞和Neuro2a细胞转染pcDNA3.1-HA-BAT3后,随着外源全长BAT3表达量的增加,内源朊蛋白的表达量表现为剂量依赖性增加。表明BAT3能与朊蛋白发生相互作用,过表达BAT3能促进内源朊蛋白的表达,提示BAT3可能影响朊蛋白的合成。To investigate the relationship between BAT3 and prion protein in vitro and explore the possible function of BAT3 in the expression of prion protein and associated diseases.The overexpression plasmid PGEFP-N1-PRNP was constructed and transfected into Hela cells and primary cortical neurons.The immunofluorescence microscopy was chosen to analyze the relationship between BAT3 and prion protein.pcDNA3.1-HA-BAT3 was constructed and transfected into Hela cells and Neuro2 a cells in different amounts.Western blotting was used to examine the expression levels of prion protein after the transfection with different doses of BAT3.BAT3 and prion protein co-localized in the cytoplasm of Hela cells and primary neuronal cells.With the increasing expression of exogenous BAT3,the endogenous prion protein levels were enhanced in Hela cells and Neuro2 a cells.Conclusion BAT3 interacted with prion protein in cytoplasm and markedly increased the endogenous expression of prion protein in Hela and Neuro2 a cells.It is possible that BAT3 might stabilize prion protein and influence its subsequent cellular function（s）.

关 键 词：BAT3 朊蛋白 原代神经元 蛋白合成 传染性海绵状脑病 

分 类 号：S852.659.7[农业科学—基础兽医学]