親和短肽用于快速檢測(cè)轉(zhuǎn)基因蛋白Cry3Bb
發(fā)布時(shí)間:2018-03-22 13:02
本文選題:CryBb毒素 切入點(diǎn):轉(zhuǎn)基因植物 出處:《江蘇農(nóng)業(yè)學(xué)報(bào)》2017年03期 論文類型:期刊論文
【摘要】:本研究利用競(jìng)爭(zhēng)洗脫法淘選與Cry3Bb毒素具有親和力的特異性短肽,以此建立間接競(jìng)爭(zhēng)酶聯(lián)免疫吸附檢測(cè)方法(IC-ELISA)。經(jīng)過(guò)4輪淘選、富集,獲得與靶蛋白具有較高親和力的B10克隆(ACIHSPTALCGGG)。所建立的檢測(cè)方法穩(wěn)定性較好,變異系數(shù)在5%以內(nèi);線性檢測(cè)范圍為0.03~29.95μg/ml,線性回歸方程為(Y=20.044x+50.407,R2=0.983 4),檢測(cè)限為0.009 6μg/ml,在玉米樣品中提取回收率為92.26%~101.22%。本研究所建立的ELISA檢測(cè)方法為糧食中Cry3Bb蛋白的定量檢測(cè)提供了有效的手段,在轉(zhuǎn)基因產(chǎn)品的檢驗(yàn)檢疫中有較高的應(yīng)用價(jià)值。
[Abstract]:In this study, an indirect competitive enzyme linked immunosorbent assay (IC-ELISAA) was established for the detection of short peptides with affinity to Cry3Bb toxin by competitive elution method. B10 clone with high affinity to target protein was obtained. The stability of the method was good, and the coefficient of variation was less than 5%. The linear detection range was 0.03 渭 g / ml, the linear regression equation was 20.044x 50.407R20.9834, the detection limit was 0.009 渭 g / ml, and the recovery rate was 92.26g / ml. The ELISA method established in this study provided an effective method for the quantitative detection of Cry3Bb protein in grain. It has high application value in the inspection and quarantine of transgenic products.
【作者單位】: 金陵科技學(xué)院園藝學(xué)院;江蘇省農(nóng)業(yè)科學(xué)院食品質(zhì)量安全與檢測(cè)研究所/農(nóng)業(yè)部食品質(zhì)量安全監(jiān)控重點(diǎn)開放實(shí)驗(yàn)室;
【基金】:江蘇省自然科學(xué)基金青年基金項(xiàng)目(BK20150114) 江蘇省高校自然科學(xué)研究項(xiàng)目(14KJB 210004) 金陵科技學(xué)院博士啟動(dòng)項(xiàng)目(jit-6-201518);金陵科技學(xué)院校級(jí)科研基金孵化項(xiàng)目(C140501)
【分類號(hào)】:S188
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本文編號(hào):1648761
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