东亚飞蝗pSMC重组抗原表达及多克隆抗体制备  

Recombinant Expression and Polyclonal Antibody Preparation of pSMC Based on the Structural Maintenance of Chromosome Protein of Locusta Migratoria

作　　者：王麒霖 邱佳 余瑛 夏玉先 WANG Qilin;QIU Jia;YU Ying;XIA Yuxian(College of Pharmacy and Biological Engineering,Chongqing University of Technology,Chongqing 400054,China;School of Life Sciences,Chongqing University,Chongqing 400030,China)

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]重庆大学生命科学学院,重庆400030

出　　处：《重庆理工大学学报（自然科学）》2021年第10期257-263,共7页Journal of Chongqing University of Technology：Natural Science

摘　　要：采用生物信息学方法对pSMC抗原特性进行分析;利用DNA重组技术构建pSMC_(249-383)重组表达菌株;以Ni亲和层析技术纯化重组pSMC_(249-383)蛋白;用重组pSMC_(249-383)蛋白免疫小鼠,制备Anti-pSMC多克隆抗体;采用免疫荧光法检测pSMC在血细胞中的定位。将pSMC编码区249-383区段插入pET30a(+),成功构建了pSMC_(249-383)原核表达菌株;用表达、纯化获得的重组pSMC_(249-383)蛋白免疫小鼠成功获得了高效价的Anti-pSMC多克隆抗体;免疫荧光检测显示pSMC在东亚飞蝗血淋巴吞噬细胞高表达,定位于细胞核,为研究pSMC功能奠定了基础。Taking the Bioinformatics methods to analyze the antigenic characteristics of pSMC;Recombinant DNA technology was used to construct pSMC_(249-383) recombinant expression strain;and the pSMC_(249-383) protein was purified by Ni affinity chromatography;Immuning mice with recombinant pSMC_(249-383) protein to produce Anti-pSMC polyclonal antibody and then detected its localization in blood cells via immunofluorescence method.The pSMC_(249-383) prokaryotic expression strain was successfully constructed by means of inserting the 249-383 region of pSMC into pET30a(+);The recombinant pSMC_(249-383) protein obtained by expression and purification was used to immunize mice and successfully generated high titer Anti-pSMC polyclonal antibody;The immunofluorescence detection results indicate that pSMC was highly expressed in the hemolymphphagocytes of Migratory locust and localized in the nucleus,thus laying a foundation for the study of pSMC function.

关 键 词：东亚飞蝗 染色体结构维持蛋白 DNA重组 生物信息学 多克隆抗体 

分 类 号：Q784[生物学—分子生物学]