巴西橡胶树肌动蛋白的原核表达及鉴定  

Prokaryotic Expression and Identification of Actin Protein in Laticifer Cells of Rubber Tree (Hevea brasiliensis Müll. Arg.)

作　　者：邓顺楠[1,2] 史敏晶[2] 吴绍华[2] 田维敏[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院橡胶研究所农业部橡胶树生物学重点开放实验室省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,海南儋州571737

出　　处：《热带作物学报》2013年第5期795-799,共5页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.30170535);现代农业产业技术体系建设专项(No.CARS-34-GW1)

摘　　要：肌动蛋白细胞骨架是橡胶树乳管伤口堵塞物的成分之一。为了研究乳管伤口末端与肌动蛋白互作的蛋白,成功构建了ACTIN基因的原核表达载体。在37℃条件下,经0.1 mmol/L IPTG诱导2 h,在E.coil BL21(DE3)中大量表达重组蛋白。该重组蛋白主要以包涵体形式存在,分子量41.7 ku。通过Ni-NTA亲和层析柱纯化了该重组蛋白,并用抗HIS标签的鼠单克隆抗体对纯化蛋白进行鉴定。本研究为揭示乳管伤口末端堵塞物形成机制打下了良好基础。ACTIN, a component of cytoskeleton, is one of the constituents of the plugs at the end of the severed laticifers. In order to study the interaction of ACTIN with proteins in the plugs, the A CTIN gene prokaryotic expression vector was successfully constructed. Recombinant protein was expressed in abundance in the E.coil BL21 （DE3） after being induced by 0.1 mmol/L IPTG for 2 hours at 37 ℃. The recombinant protein with a molecular mass of about 41.7 ku was mainly present as inclusion body. It was further purified by Ni-NTA affinity chromatography and identified by western-blot with mouse anti-HIS tagged monoclonal antibody. This study laid a good foundation of revealing the mechanism for plug formation at the end of severed laticifers.

关 键 词：巴西橡胶树 肌动蛋白细胞骨架 乳管堵塞 原核表达 

分 类 号：S794.1[农业科学—林木遗传育种]