Overexpression and purification of an immunologically reactive His-BIV capsid fusion protein.
Overexpression and purification of an immunologically reactive His-BIV capsid fusion protein.
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免疫反应性 His-BIV 衣壳融合蛋白的过表达和纯化。
DOI：
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发表时间：
1999
影响因子：
1.6
通讯作者：
T. Baron
中科院分区：
文献类型：
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作者：
D. Bétemps;F. Mallet;V. Cheynet;T. Baron
将牛免疫缺陷病毒（BIV）衣壳蛋白的基因与编码六个组氨酸的序列连接并表达为（His）6 p26衣壳融合蛋白。在异丙硫基-β-d-半乳糖苷诱导后，融合蛋白以可溶性和不溶性形式强烈表达。纯化基于六组氨酸多肽与金属离子的相互作用。表达量可占大肠杆菌总蛋白的 11%，每升细菌培养物可获得超过 20 毫克的高纯度蛋白。通过固定化金属亲和层析纯化的 (His)6 p26 衣壳融合蛋白在蛋白质印迹中与实验性感染 BIV 的牛血清以及针对 Gag 蛋白不同表位的两种单克隆抗体发生特异性反应。这种融合蛋白易于表达、纯化和特异性，应该允许在现场样本的大规模血清学研究中彻底研究 BIV 感染的流行情况。
The gene of the capsid protein of bovine immunodeficiency virus (BIV) was linked to a sequence encoding for six histidines and expressed as the (His)6 p26 capsid fusion protein. The fusion protein was strongly expressed as both soluble and insoluble forms after induction by isopropylthio-beta-d-galactoside. Purification was based on interaction of the hexa-histidine polypeptide with metal ions. Expression could represent 11% of the total protein in Escherichia coli, allowing more than 20 mg of highly purified protein to be obtained per liter of bacterial culture. The (His)6 p26 capsid fusion protein purified by immobilized metal affinity chromatography reacted specifically in Western blot with sera from cattle experimentally infected by BIV, as well as with two monoclonal antibodies directed against different epitopes of the Gag protein. The ease of expression, purification, and specificity of this fusion protein should permit a thorough study of prevalence of BIV infection in large-scale serological studies of field samples.
影响因子：
3.7
作者：
D. Suarez;C. Whetstone
通讯作者：
D. Suarez;C. Whetstone
使用细菌 trpE 融合载体表达和表征牛免疫缺陷样病毒核心蛋白。
影响因子：
3.1
作者：
Atkinson,B;Liu,ZQ;Wood,C
通讯作者：
Wood,C
使用 TrpE 融合蛋白识别 BIV 包膜蛋白内的抗原结构域。
影响因子：
3.1
作者：
Chen,P;Liu,ZQ;Wood,C
通讯作者：
Wood,C