神经系统富亮氨酸重复超家族成员LRRN3 C端结构域蛋白的原核表达和纯化  

Expression and Purification of C-Terminal Domain in LRRN3,a Member of Neuron Leucine Rich Repeat Superfamily

作　　者：杨静[1] 蔡艳[2] 黄河[1] 李芳[2] 伍赶球[1] 杨景[1] 罗学港 

机构地区：[1]中南大学湘雅医学院组织学与胚胎学系,中国湖南长沙410013 [2]中南大学湘雅医学院人体解剖学与神经生物系,中国湖南长沙410013

出　　处：《生命科学研究》2008年第3期227-231,共5页Life Science Research

基　　金：国家自然科学基金资助项目(30600224;30700438);中国博士后科学基金资助项目(20060390886);湖南省自然科学基金资助项目(06JJ30014)

摘　　要：神经系统富亮氨酸重复超家族成员LRRN3是一种重要的膜蛋白,与神经系统发生发育和损伤后修复密切相关.运用多聚酶链式反应(PCR)方法,获得长555bp的DNA序列,扩增产物克隆至pET21质粒中,构建Mal和His融合表达质粒,在大肠杆菌中诱导表达融合蛋白,经Ni+-NTA agarose亲和层析纯化获得融合蛋白,并以Western blotting鉴定,为进一步研究LRRN3基因的结构和功能奠定了基础.DNA sequence of C-terminal domain of LRRN3, a member of neuron leucine rich repeat superfamily, was obtained by PCR. A 555 bp sequence was cloned into expression vector pET21 to construct the Mal and His fusion protein. The recombinant plasmid was transformed into E. coli BL21 and the over expressed fusion protein of C-terminal domain was existed in inclusion bodies. The fusion protein was purified with Ni^＋ -NTA agarose. Fusion protein was testified by Western blotting using anti-His monoelonal antibody. The foundation was based by this work to do further investigation on the structure and function of LRRN3.

关 键 词：LRRN3 融合蛋白 蛋白纯化 

分 类 号：Q786[生物学—分子生物学]