兔抗仿蜘蛛拖丝蛋白多克隆抗体的制备与应用

Ting-Qing Guo, Yun Zhao, Sheng-Peng Wang, Liang Dong, Chang-De Lu
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引用次数: 0

摘要

将合成蜘蛛拖丝基因s600克隆到融合蛋白表达载体pGEX-KG中,并在大肠杆菌中表达。纯化了S600蛋白,制备了兔抗血清。氨基酸组成分析证实S600正确表达。免疫印迹分析表明,抗s600抗血清能与天然蜘蛛丝发生反应,因此设计的合成拖丝蛋白具有与天然蜘蛛丝相似的免疫特性。建立了转基因家蚕蚕丝腺(或蚕茧)中合成拖丝蛋白表达的ELISA定量检测系统。
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[Production and application of rabbit anti-imitative spider dragline silk protein polyclonal antibody].

A synthetic spider dragline gene s600 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.

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