家蚕丝素轻链蛋白多克隆抗体的制备及应用  

Preparation and Application of Silkworm Fibroin Light Chain Polyclone Antibody

作　　者：刘春[1] 韩宇[1] 荣海博[1] 侯勇[1] 陈玉琳[1] 高志宏[1] 夏庆友[1] 

机构地区：[1]西南大学蚕学与系统生物学研究所,重庆400716

出　　处：《蚕学通讯》2009年第1期9-14,共6页Newsletter of Sericultural Science

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB121000);国家"863"计划项目(2006AA10A118)

摘　　要：本研究通过用回收纯化的Fib-L蛋白为抗原免疫家兔,获得多克隆抗体,通过与家蚕大造5龄5d的10个不同组织进行Western blot,结果表明,制备的Fib-L抗体能识别在后部丝腺特异合成的Fib-L蛋白,表明该抗体有较好的特异性。利用此抗体进行丝腺组织的免疫组化,经抗体的一系列的梯度稀释和条件探索,发现当Fib-L一抗在1∶50和荧光二抗1∶1000的条件下得到理想的免疫组化结果,在荧光显微镜下观察到了二抗标记的绿色荧光。以上结果显示,本实验成功制得Fib-L蛋白抗体,获得丝腺组织免疫组化的一些重要参考信息,为家蚕丝腺功能基因的研究及细胞定位提供了一定的参考价值。To obtain polyclone antibody against Fib-L, rabbit was immunized by multiple-site hy- podermic injection with the purified Fib-L proteins. The specificity of the antibody was verified with Western blot assay against proteins from 10 different tissues of silkworm Dazao, and the result showed that the antibody could hybridize with Fib-L proteins only in the posterior silk gland. With the antibody, assay of immunohistochemistry with silk gland was per- formed. Under the conditions of Fib-L antibody with 1.50 diluted and fluorescence anti-IgG with1 ： 1000, green fluorescence was detected with the fluorescence microscope. The successful preparation of the antibody against Fib-L is an elementary exploration to the production of polyclone antibody. Meanwhile, to some extent, it is a basis for the study of immunohistochemistry, silk gland function gene research and subcellular localization.

关 键 词：丝素基因 Fib-L蛋白 多克隆抗体 WESTERN blot免疫组化 

分 类 号：S881[农业科学—特种经济动物饲养]