Copious production of SARS-CoV nucleocapsid protein employing codon optimized synthetic gene.
Copious production of SARS-CoV nucleocapsid protein employing codon optimized synthetic gene.
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DOI：
10.1016/j.jviromet.2006.06.029
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发表时间：
2006-11
影响因子：
3.1
通讯作者：
Suresh MR
中科院分区：
文献类型：
--
作者：
Das D;Suresh MR
严重急性呼吸综合征冠状病毒（SARS CoV）核衣壳蛋白（NP）是SARS病毒感染过程中最主要的抗原蛋白之一，也是最丰富的脱落抗原。该特征使其成为诊断应用的合适分子靶标。本研究将密码子优化的NP基因及其亚片段克隆到细菌表达载体中。在E.大肠杆菌中的包涵体中有很高的表达水平。包涵体成功地溶解，在变性条件下使用IMAC柱纯化并复性。非糖基化NP用于免疫小鼠用于杂交瘤开发。用该重组NP蛋白免疫的动物的多克隆抗血清被发现特异性地识别NP及其亚片段，从而证明了重组蛋白的免疫原性。NP抗原或其亚片段可用于建立一种灵敏的血清学诊断方法，通过检测早期人抗SARS抗体来监测SARS冠状病毒的爆发。此外，NP片段的可用性可以促进抗NP单克隆抗体的表位作图，以鉴定合适的夹心对。
The severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein (NP) is one of the predominant antigenic protein and the most abundant shed antigen throughout the SARS-CoV infection. This feature makes it a suitable molecular target for diagnostic applications. In this study the full length codon optimized NP gene and its subfragment gene segment was cloned in a bacterial expression vector. The full length NP could be expressed in E. coli at very high level within inclusion bodies. The inclusion bodies were successfully solubilized, purified under denaturing conditions employing IMAC column and refolded. The non-glycosylated NP was used to immunize mice for hybridoma development. The polyclonal antiserum from animals immunized with this recombinant NP protein was found to specifically recognize the NP and its subfragments, thus demonstrating the immunogenic nature of the recombinant protein. The NP antigen or a subfragment could be useful for developing a sensitive serum diagnostic assay to monitor SARS-CoV outbreaks by detecting the early human anti-SARS antibodies. In addition, the availability of the NP fragments could facilitate epitope mapping of anti-NP monoclonals for identifying suitable sandwich pairs.
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影响因子：
5
作者：
HISCOX, JA;CAVANAGH, D;BRITTON, P
通讯作者：
BRITTON, P
影响因子：
11.8
作者：
Chan PK;Ng KC;Chan RC;Lam RK;Chow VC;Hui M;Wu A;Lee N;Yap FH;Cheng FW;Sung JJ;Tam JS
通讯作者：
Tam JS
DOI：
10.1128/cdli.11.2.417-422.2004
发表时间：
2004-03-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
He, QG;Chong, KH;Kwang, J
通讯作者：
Kwang, J
影响因子：
9.4
作者：
Che, XY;Qiu, LW;Yuen, KY
通讯作者：
Yuen, KY
影响因子：
4.1
作者：
SOHN, MJ;CHONG, YH;LEE, YI
通讯作者：
LEE, YI