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

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.