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[兔抗仿蜘蛛拖牵丝蛋白多克隆抗体的制备及应用]

[Production and application of rabbit anti-imitative spider dragline silk protein polyclonal antibody].

作者信息

Guo Ting-Qing, Zhao Yun, Wang Sheng-Peng, Dong Liang, Lu Chang-De

机构信息

Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China.

出版信息

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Aug;35(8):756-60.

Abstract

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.

摘要

将合成的蜘蛛拖牵丝基因s600克隆到融合蛋白表达载体pGEX-KG中,并在大肠杆菌中表达。对蛋白S600进行纯化并制备兔抗血清。氨基酸组成分析证实了S600的正确表达。蛋白质免疫印迹分析表明,抗S600抗血清可与天然蜘蛛丝发生反应,因此作者设计的合成拖牵丝蛋白与天然蜘蛛丝具有相似的免疫学特性。还建立了一种酶联免疫吸附测定(ELISA)系统,用于定量检测转基因家蚕丝腺(或茧中)合成拖牵丝蛋白的表达。

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