果蝇血液发育标记基因mrj的多克隆抗体制备及检测  

Preparation and Assay of Polyclonal Antibody of Blood Development Marker Gene Mrj in Drosophila Melanogaster

作　　者：雷旻音[1] 杨哲[1] 黄婷[1] 刘丹[1] 李永青[1] 吴秀山[1] 

机构地区：[1]湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室,心脏发育研究中心,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2012年第4期55-58,共4页Journal of Natural Science of Hunan Normal University

基　　金：国家自然科学基金资助项目(30671053;30871340;30930054)

摘　　要：mrj是黑腹果蝇中一个蛋白编码基因.根据已报道的mrj基因序列,利用生物信息学分析选取果蝇mrj基因抗原亲水区,通过PCR扩增出其部分编码区序列,将其连接到pET-28a原核表达载体上.经酶切及测序鉴定质粒构建成功后,将重组质粒(pET-28a-mrj)转入Rosetta菌后通过IPTG(Isopropy1β-D-thiogalactoside)诱导表达出带His标签的重组融合蛋白后用镍柱纯化.将纯化的蛋白免疫新西兰大白兔制备多克隆抗体,并用Western Blot检测抗体的效价和特异性.Mrj is a protein coding gene from Drosophila. Hydrophilic and well specific fragment of mrj is selected by bioinformatics method. Then it was amplified the fragment by PCR and cloned into the expression vector pET-28a to acquire the recombinant expression plasmid pET-28a-mrj. It was transformed into Escherichia coli Rosetta, and the fusion protein with His tag was induced with IPTG. The fusion protein was purified by nickel chelating resin. His-mrj fusion protein was used to immune the New Zealand white rabbits, then identified the antibody by Western blot.

关 键 词：mrj基因 果蝇 多克隆抗体 

分 类 号：Q78[生物学—分子生物学]