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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
【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 ,建立了用ELISA方法定量检测茧壳中仿蜘蛛牵丝蛋白量的工作系统
【Abstract】 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.
【Key words】 synthetic spider dragline; rabbit anti serum; cocoon; ELISA;
- 【文献出处】 生物化学与生物物理学报 ,Acta Biochimica Et Biophysica Sinica , 编辑部邮箱 ,2003年08期
- 【分类号】Q78
- 【被引频次】6
- 【下载频次】117