| 斑节对虾酚氧化酶原在大肠杆菌中的表达 |
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| 引用本文： | 韩凤丽,贡成良,曹广力,薛仁宇.斑节对虾酚氧化酶原在大肠杆菌中的表达[J].常熟理工学院学报,2009,23(4):61-64. |
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| 作者姓名： | 韩凤丽 贡成良 曹广力 薛仁宇 |
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| 作者单位： | 苏州大学医学部,江苏苏州,215123 |
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| 摘 要： | 将斑节对虾酚氧化酶原（prophenoloxidase，proPO）基因克隆进pET28a（＋），在大肠杆菌BL21菌株中诱导表达，SDS-PAGE和Western-blotting分析均能检测到一条分子量为79．4kDa的特异性条带，与推导的融合蛋白理论分子量82．4kDa基本相符；包涵体变性后对经Ni—NTAagarose纯化的变性液进行复性，表现出0．3851U／mg．min的PO活性；不同条件下的诱导表达结果显示，18℃时诱导培养能检测到目的蛋白的可溶性表达，经诱导9h后可溶性表达比例达到最高，约占菌体可溶性蛋白总量的s．56％；可溶性表达的目的蛋白纯化后经胰酶消化，可检测到分子量为60kDa、42．7kDa的特异性条带，并表现出0．4249U／mg．min的PO活性．
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| 关 键 词： | 蛋白纯化 proPO 原核表达 酶活性 |
Expression and Purification of Prophenoloxidase of the Black Tiger Shrimp(Penaurs Monodon)in E.coli |
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| HAN Feng-Li,GONG Cheng-Liang,CAO Guang-Li,XUE Ren-Yu.Expression and Purification of Prophenoloxidase of the Black Tiger Shrimp(Penaurs Monodon)in E.coli[J].Journal of Changshu Institute of Technology,2009,23(4):61-64. |
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| Authors: | HAN Feng-Li GONG Cheng-Liang CAO Guang-Li XUE Ren-Yu |
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| Institution: | College of Medicine;Suzhou University;Suzhou 215123;China |
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| Abstract: | The prophenoloxidase gene of the black tiger shrimp(Penaurs monodon) was cloned into pET28a(+) vector and expressed in E.coli BL21.A special band of 79.4kDa was detected using SDS-PAGE and Western blot, which matched the theoretical molecular mass(82.4kDa) of the fusion protein on the whole.The obtained inclusion body was denatured,purified with nickel-resin column and renatured,resulting in the enzyme activity of 0.3851 U/ mg.min.The transformed E.coli BL21 were induced under different conditions.The solub... |
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| Keywords: | proPO |
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