兔抗仿蜘蛛牵丝蛋白抗体的制备及应用  

Production and Application of Rabbit Anti-imitative Spider Dragline Silk Protein Polyclonal Antibody

作　　者：郭廷晴[1] 赵昀[1] 汪生鹏[1] 董良[1] 陆长德[1] 

机构地区：[1]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《生物化学与生物物理学报》2003年第8期756-760,共5页

基　　金：上海市现代生物与医药产业发展基金(No .0 2 43 192 0 9);中国科学院上海生命科学研究院研究发展基地资助项目~~

摘　　要：将仿蜘蛛牵丝基因s6 0 0克隆到GST融合蛋白表达质粒pGEX KG中 ,利用大肠杆菌表达系统表达并纯化了仿蜘蛛牵丝蛋白S6 0 0 ,以之作为抗原制备了兔抗血清。S6 0 0的氨基酸组分分析与理论值相吻合。蛋白质免疫印迹发现该抗血清能与天然蜘蛛丝反应 ,表明设计的仿蜘蛛丝与天然蜘蛛丝有相似的免疫原性。为了定量检测仿蜘蛛牵丝蛋白在转基因家蚕丝腺中 (或茧壳中 )的表达 。A synthetic spider dragline gene s 600 was cloned into fusion protein expression vector pGEX KG and expressed in Escherichia coli. Protein S600 was purified and rabbit antiserum was prepared. Amino acid composition analysis confirmed the right expression of S600. Western blot analysis revealed that anti S600 antiserum could react with natural spider silk, so the synthetic dragline protein, designed by the authors, shares similar immunological characteristics with the natural spider silk. An ELISA system was also established for the quantitative detection of synthetic dragline protein expression in silk gland (or in the cocoon) of transgenic silkworm.

关 键 词：仿蜘蛛牵丝蛋白 兔抗血清 茧壳 抗体 蛋白质免疫印迹 

分 类 号：Q78[生物学—分子生物学]