集胞藻6803藻胆体藻蓝蛋白多克隆抗体制备及其初步应用  

Preparation and preliminary application of polyclonal antibody of c-phycocyanin protein in the cyanobacterium Synechocystis sp. strain PCC 6803

作　　者：张劲松[1] 陈李萍[1] 高复旦[1] 杜玲瑜[1] 王全喜[1] 马为民[1] 

机构地区：[1]上海师范大学生命与环境科学学院,上海200234

出　　处：《广东农业科学》2010年第11期1-5,9,共6页Guangdong Agricultural Sciences

基　　金：国家自然科学基金(30770175);国家"973"计划项目(2009CB118500);教育部重点项目(209045);上海市教委科研创新重点项目(08ZZ67)

摘　　要：通过PCR扩增出集胞藻6803 C-PC编码基因cpcA,构建表达质粒pET32a(+)-cpcA,转化大肠菌株BL21(DE3)pLysS,用IPTG诱导表达、经His-tag纯化后免疫日本大耳白兔获得多克隆抗体。间接ELISA法揭示该抗体效价可高达1∶1 025 000;蛋白免疫印迹确定该抗体具有高度特异性。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量,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。In this study,the encoding gene,cpcA,was PCR amplified from the unicellular cyanobacterium Synechocystis sp.strain PCC 6803,the expression plasmid pET32a（＋）-cpcA 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 ∶1025000,and possessed high specificity.Further,the amounts of C-PC protein were analyzed under the acclimatization of the unicellular cyanobacterium cells to growth and high light,and between two photosystems by using the polyclonal antibody.The results indicated the existence of the remarkable differences in C-PC protein amounts under these 2 situations.This will further help us to reveal the important roles of the peripheral rods of cyanobacterial phycobilisomes underlying the responding and acclimating mechanisms.

关 键 词：藻蓝蛋白 多克隆抗体 集胞藻6803 

分 类 号：Q949.2[生物学—植物学]