The expression of SARS-CoV M gene in P Pastoris and the diagnostic utility of the expression product
The expression of SARS-CoV M gene in P Pastoris and the diagnostic utility of the expression product
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DOI：
10.1016/j.jviromet.2004.08.015
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发表时间：
2004-12-01
影响因子：
3.1
通讯作者：
Rao, ZH
中科院分区：
文献类型：
--
作者：
Han, XQ;Bartlam, M;Rao, ZH
高水平蛋白表达是获得大量病毒蛋白以进一步研究其生物学特性的重要手段。为了在体外高水平表达SARS-CoV的膜（M）蛋白，扩增了M基因片段并将其克隆到毕赤酵母表达载体pPICZalphaA中。重组酵母转化子诱导产物的SDS-PAGE和Western blotting分析表明M蛋白成功高水平表达，且表达产物与天然蛋白具有相似的抗原性。随后以纯化的重组M蛋白作为ELISA抗原对之前通过全病毒ELISA和免疫荧光法筛选的8份血清样本进行检测，得到了一致的结果。这些发现表明重组 M 蛋白可用作诊断试剂。 (C) 2004 Elsevier B.V. 保留所有权利。
High-level protein expression is an important means of obtaining large amounts of viral proteins to investigate further their biological properties. To express the membrane (M) protein of SARS-CoV at high-level in vitro, the M gene fragment was amplified and cloned it into the Pichia Pastoris expression vector pPICZalphaA. SDS-PAGE and Western blotting analysis of the induced products of recombinant yeast transformant indicated that successful high-level expression of M protein was achieved, and that the expression product was similar antigenically to the natural protein. Purified recombinant M protein was used subsequently as an ELISA antigen for detection of eight serum samples screened previously by whole virus ELISA and immunofluorescence assay, and consistent results were obtained. These findings suggest that the recombinant M protein may be useful as a diagnostic reagent. (C) 2004 Elsevier B.V. All rights reserved.