GnRH缺陷疾病致病基因RNF216互作蛋白的筛选  

Screening of interacting proteins of idiopathic gonadotropin-releasing hormone deficiency pathogenic gene RNF216

作　　者：戴文婷 蒋最明 顾敏 朱勇[2] 唐满玲 陈祥 Dai Wenting;Jiang Zuiming;Gu Min;Zhu Yong;Tang Manling;Chen Xiang(Center of Clinical Laboratory,the Affiliated Zhuzhou Hospital of Xiangya Medical College,Central South University,Zhuzhou,Hunan 412007,China;Department of Blood Transfusion,the Affiliated Zhuzhou Hospital of Xiangya Medical College,Central South University,Zhuzhou,Hunan 412007,China)

机构地区：[1]中南大学湘雅医学院附属株洲医院临床检验中心,株洲412007 [2]中南大学湘雅医学院附属株洲医院输血科,株洲412007

出　　处：《中华医学遗传学杂志》2021年第7期631-634,共4页Chinese Journal of Medical Genetics

摘　　要：目的通过酵母双杂交实验筛选与环指蛋白216(ring finger protein 216,RNF216)相互作用的蛋白,进一步阐明RNF216在GnRH缺陷疾病中的作用。方法构建pGBKT7-RNF216重组表达载体,将其转化到Y2HGold酵母中,与人cDNA文库进行杂交,筛选与RNF216相互作用的蛋白,然后在Y2HGold酵母中进行验证。结果成功构建了pGBKT7-RNF216重组表达载体,并在Y2HGold酵母中成功表达;通过酵母双杂交实验,筛选到了一个与RNF216相互作用的蛋白——丝状蛋白B(filamin B,FLNB),并在Y2HGold酵母中验证了它们之间的相互作用。结论成功筛选到一个与RNF216相互作用的FLNB蛋白,RNF216可能通过调节FLNB或FLNB/FLNA异源二聚体影响GnRH神经元的增殖和迁移。Objective To screen proteins interacting with ring finger protein 216(RNF216)through yeast two hybrid experiment,and further clarify the role of RNF216 in the pathogenesis of gonadotropin-releasing hormone deficiency.Methods A recombinant expression vector pGBKT7-RNF216 was constructed and transformed into yeast Y2HGold,which was hybridized with a human cDNA library in order to screen proteins interacting with RNF216.The interaction was verified in yeast Y2HGold.Results A recombinant expression vector pGBKT7-RNF216 was successfully constructed and expressed in yeast Y2HGold.Filamin B(FLNB)was identified by yeast two hybrid experiment,and their interaction was verified in yeast Y2HGold.Conclusion An interaction between FLNB and RNF216 was identified through yeast two hybrid experiment.RNF216 may affect the proliferation and migration of GnRH neurons by regulating FLNB or FLNB/FLNA heterodimers.

关 键 词：GnRH缺陷疾病 GnRH神经元 RNF216蛋白 FLNB蛋白 

分 类 号：R596[医药卫生—内科学]