摘要
阐述人脑内硒蛋白在维护细胞正常功能、抵御氧化损伤和预防脑疾病方面的重要作用,指出脱碘酶3是人脑中一种重要的硒蛋白.克隆了人脱碘酶3的开放读码框,将其编码区中编码硒代半胱氨酸的TGA码突变为编码半胱氨酸的密码,以脱碘酶3突变体为"诱饵",利用酵母双杂交系统从人胎脑cD-NA文库中筛选一个能与脱碘酶3相互作用的蛋白,即人丝氨酸蛋白酶抑制剂A族蛋白3.采用荧光共振能量转移技术中的敏化发射法和荧光寿命法,验证了人脑中脱碘酶的相互作用.
Selenoproteins play important roles in maintaining cell function,resisting peroxidation damage,and preventing brain diseases.Iodothyronine deiodinase 3（DIO3） is an important selenoprotein in the human brain.In this study,a human DIO3 gene was cloned and mutated at the TGA codon in the open reading frame encoding for a selenocysteine（Sec） residue,where the Sec was site-mutated to a cysteine residue.The DIO3′ mutant was then used as a ＂bait＂ to screen a fetal brain cDNA library by yeast two-hybrid system.Preliminary investigation on the protein-protein interaction was performed by the life-time-resolved imaging measurement using the fluorescence resonance energy transfer technique.Results showed that the interactive protein of DIO3′ is a serpin peptidase inhibitor（clade A,member 3）.
出处
《深圳大学学报（理工版）》
EI
CAS
北大核心
2011年第5期423-429,共7页
Journal of Shenzhen University(Science and Engineering)
基金
国家自然科学基金资助项目(31070731
30901182)~~
关键词
基因工程
硒蛋白
脱碘酶3
大脑
基因编码
酵母双杂交系统
蛋白质筛选
荧光共振能量转移技术
genetic engineering
selenoprotein
iodothyronine deiodinases 3（DIO3）
brain
gene encoding
yeast two-hybrid system
screening of protein
fluorescence resonance energy transfer（FRET）
