水稻类非特异脂质转移蛋白OSDIR的表达和纯化  

Expression and Purification of A-Rice Nonspecific Lipid Transfer Protein Homologue OSDIR

作　　者：刘晓灿[1] 汪笑峰[1] 葛晓春[1] 曹凯鸣[1] 

机构地区：[1]复旦大学生命科学学院生物化学系

出　　处：《广西师范大学学报（自然科学版）》2006年第1期75-78,共4页Journal of Guangxi Normal University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30370297)

摘　　要：通过PCR方法从水稻(O ry za sa tiva)染色体基因组中扩增一个编码非特异性脂质转移蛋白(nonspec ific lip id transfer prote in,nsLTP)类似蛋白的基因(OSd ir),将该基因片段克隆到硫氧还蛋白融合表达载体pET 32a(+)中后,在BL 21(DE 3)trxB-宿主菌中实现了融合蛋白的高表达,表达产物为包涵体。利用尿素溶解包涵体并复性后,再通过Sepharose N i2+-che lating fast flow柱亲和纯化融合蛋白,得到重组O SD IR蛋白。A gene fragment encoding nonspecific lipid transfer protein （nsLTP）-like protein OSDIR was amplified from rice （Oryza sativa L. ssp. indica） genomic DNA by PCR. This gene was cloned into Thio- FusionTM expression vector pET32a（＋） and expressed in host strain BL21 （DE3） trxB-. After induction by IPTG at 30℃ for 5 h,the fusion protein Thio-OSDIR was produced abundantly as inclusion body. After dissolved in denature reagent and renatured,the fusion protein was purified by Ni^2＋ -chelating Sepharose fast flow column. The phylogenetic relationship of OSDIR with other nsLTP members in rice was discussed.

关 键 词：生物化学 非特异性脂质转移蛋白 构建 表达 纯化 

分 类 号：Q786[生物学—分子生物学]