利用SPR原理研究mGAT-1基因5’区域和组织核蛋白的相互作用  

STUDY ON THE INTERACTION BETWEEN THE 5' PROXIMAL REGION OF mGAT-1 AND NUCLEAR PROTEINS BY THE METHOD OF SPR

作　　者：王琼[1] 蔡国强[1] 费俭[1] 周美云[1] 郭礼和[1] 

机构地区：[1]中科院上海细胞生物学研究所,上海200031

出　　处：《实验生物学报》1999年第3期221-225,共5页Acta Biologiae Experimentalis Sinica

基　　金：科学院基础局重点课题基金;上海细胞所细胞生物学开放实验室课题

摘　　要：用PCR方法扩增到小鼠GAT-1基因5’近侧序列-1775—-1594片段(F182DNA),通过生物素偶联到感应片(SA5)上,利用生物传感器,研究了小鼠肾、肝组织核蛋白提取液和F182DNA的相互作用,实验表明不同浓度的小鼠肾、肝核蛋白与F182DNA片段结合明显,并且它们的表观解离速率常数都是k_d=1.4E-5/秒。实验还发现,在F182序列中一人鼠保守的序列是一个蛋白主要的结合位点,证明这一保守序列在mGAT-1基因调控中可能起重要的作用。The DNA fragment (named F182) corresponding the position of -1775-1594 in the mouse GABA transporter 1 (mGAT-1) 5' proximal region was amplified by PCR. Then the DNA was immobilized to the surface of sensor chip SA5 via biotin-streptavidin linkage. The interaction between the F182 on SA5 and nuclear proteins from mouse liver and kidney was studied by the method of SPR with Biosensor of BIAcore-1000 respectively. The Binding between F182 and two nuclear proteins was definitely and specifically and both with the apparent dissociation rate of about 1. 4E-5/S. Competitive experiment revealed that a conserved sequence within F182 had the main contribution to the binding event.

关 键 词：mGAT-1 组织核蛋白 转运蛋白 神经递质 基因调控 

分 类 号：Q422[生物学—神经生物学] Q756[生物学—生理学]