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Genetic Analysis and Expression of NS3 Gene of Bovine Viral Diarrhoea Virus 1 from India for Detection of Antibodies in Cattle

机译:印度牛腹泻病毒1号NS3基因的遗传分析和表达以检测牛的抗体

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Mishra, N., Pitale, S.S. and Pradhan, H.K. 2008. Genetic analysis and expression of NS3 gene of bovine viral diarrhoea virus 1 from India for detection of antibodies in cattle. J. Appl. Anim. Res., 33: 99?¢????103. Considering the importance of NS3 antigen in diagnosis of bovine viral diarrhea virus (BVDV) infection, we analysed genetically selected Indian isolates in NS3 gene region and generated recombinant NS3 protein in Escherichia coli for its use as ELISA antigen. High degree of conservation was observed when the nucleotide and deduced amino acid sequence of Indian and the reference BVDV 1 isolates were analysed. The helicase domain of NS3 gene of an Indian isolate was cloned into pTriEx-2-Neo expression vector and a recombinant protein of 50 KDa was expressed as cytoplasmic inclusion bodies. The affinity purified protein showed antigenic properties in western blot with BVDV infected cattle serum. When used in ELISA, it could detect and BVDV antibodies in bovine sera. The expression of NS3 recombinant protein showing immunological and diagnostic significance is first such study in India.
机译:N.Mishra,S.S。Pitale和H.K. Pradhan 2008。来自印度的牛病毒性腹泻病毒1的NS3基因的遗传分析和表达,用于检测牛中的抗体。 J.应用动画。 Res.33:99-103。考虑到NS3抗原在诊断牛病毒性腹泻病毒(BVDV)感染中的重要性,我们分析了NS3基因区域中遗传选择的印度分离株,并在大肠杆菌中产生了重组NS3蛋白作为ELISA抗原。当分析Indian和参照BVDV 1分离株的核苷酸和推导的氨基酸序列时,观察到高度的保守性。将印度分离株的NS3基因的解旋酶结构域克隆到pTriEx-2-Neo表达载体中,并将​​50KDa的重组蛋白表达为细胞质包涵体。亲和纯化的蛋白在BVDV感染的牛血清中进行Western印迹显示抗原性。当用于ELISA时,它可以检测和检测牛血清中的BVDV抗体。具有免疫学和诊断意义的NS3重组蛋白的表达是印度的首次此类研究。

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