本文選題：P　切入點(diǎn)：CYPJ融合蛋白　出處：《生物技術(shù)通報(bào)》2017年05期 　論文類型：期刊論文

【摘要】：通過親和層析法純化棉蚜His-CYP6J1融合蛋白,制備及鑒定其多克隆抗體。采用鎳離子金屬螯合親和層析柱分離純化棉蚜His-CYP6J1融合蛋白,SDS-PAGE檢測純化產(chǎn)物。用純化得到的His-CYP6J1融合蛋白免疫小鼠,制備抗棉蚜P450CYP6J1多克隆抗體;采用ELISA法檢測抗體效價;免疫組化法檢測抗血清特異性。結(jié)果表明,純化的His-CYP6J1融合蛋白免疫小鼠后得到多抗血清,ELISA法檢測抗血清效價達(dá)1∶200 000。免疫組化結(jié)果顯示,此多克隆抗體能夠與棉蚜P450 CYP6J1天然蛋白特異性結(jié)合,卻在棉鈴蟲沒有發(fā)現(xiàn)相應(yīng)的結(jié)合反應(yīng)。上述結(jié)果對研究棉蚜單一P450蛋白結(jié)構(gòu)、功能及其在棉蚜抗藥性形成過程的作用奠定基礎(chǔ)。
[Abstract]:His-CYP6J1 fusion protein of cotton aphid was purified by affinity chromatography. The polyclonal antibody was prepared and identified. The purified product of His-CYP6J1 fusion protein of cotton aphid (APHIS gossypii) was separated and purified by nickel ion metal chelate affinity chromatography. The purified His-CYP6J1 fusion protein was used to immunize mice to prepare the polyclonal antibody against APHIS gossypii P450 CYP6J1. The antibody titer was detected by ELISA method, and the specificity of antiserum was detected by immunohistochemical method. The results showed that after immunizing mice with purified His-CYP6J1 fusion protein, the titer of antiserum was 1: 200000 by Elisa. The polyclonal antibody could specifically bind to the natural protein P450 CYP6J1 of cotton aphid, but no corresponding binding reaction was found in cotton bollworm. Function and its role in the formation of resistance to cotton aphids laid the foundation.
【作者單位】： 新疆大學(xué)生命科學(xué)與技術(shù)學(xué)院新疆生物資源基因工程重點(diǎn)實(shí)驗(yàn)室;
【基金】：自治區(qū)青年科技創(chuàng)新人才培養(yǎng)工程項(xiàng)目(qn2015yx001) NSFC-新疆聯(lián)合項(xiàng)目(U1603331)
【分類號】：S433.39

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