集胞藻6803 藻胆体藻蓝蛋白多克隆抗体制备及其初步应用.pdfVIP

广东农业科学 20 10 年第11 期 1 胞藻6803 藻胆体藻蓝蛋白多克隆 抗体制备及其初步应用 劲松 陈李萍 高复旦 杜玲瑜 王全喜 马为民 ( 海师范大学生命与环境科学学院 上海 200234) 摘要院通过PCR 扩增出集胞藻6803 C-PC 编码基因cpc A 构建表达质粒pET32a(+)-cpc A 转化大肠菌株BL21(DE3) pLysS 用IPTG 诱导表达尧经His-tag 纯化后免疫日本大耳白兔获得多克隆抗体遥 间接ELISA 法揭示该抗体效价可高达 1颐 1 025 000 曰蛋白免疫印迹确定该抗体具有高度特异性遥 应用该抗体进行蛋白免疫印迹检测 结果发现C-PC 蛋白在生长光 与高光培养条件下的数 以及与2 个光系统间的连结数量均存在显著差异 为进一步研究藻胆体的杆在响应与适应机 制中所承担的重要角色奠定了生化基础遥 关键词院藻蓝蛋白曰 多克隆抗体曰集胞藻6803 中图分类号院Q949.2 文献标识码院A 文章编号院1004-874X 渊2010冤11-0001-05 Preparation and preliminary application of polyclonal antibody of c-phycocyanin protein in the cyanobacterium Sy nechocystis sp. strain PCC 6803 ZHANG Jin-song, CHEN Li-ping, GAO Fu-dan, DU Ling-yu, WANG Quan-xi, MA Wei-min (College of L ife and Env ironment Sc iences, Shanghai Normal Univ ers ity, Shanghai 200234, China) Abstract 院 In this study, the encoding gene, cpc A, was PCR amplified from the unicellular cyanobacterium Synechocystis sp. strain PCC 6803, the expression plasmid pET32a (+)-cpc A was constructed and transformed into BL21 (DE3)pLysS, and the expression of C -PC protein was induced by IPTG. After purification, the fusion protein His -C -PC was used to immunize Japanese white rabbit to obtain the polyclonal antibody. The titer of the polyclonal antibody was detected by ELISA and its specificity was analyzed by immunoblotting. The results indicated that the titer of polyclonal antibody was 1 颐1025000, and possessed high specificity. Further, the amounts of C -PC protein were anal