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Prokaryotic expression and polyclonal antibody preparation of novel ZLG10 protein involved in infection of RSV on SPC-A1 cells

机译:新型RSG10蛋白参与SPC-A1细胞RSV感染的原核表达及多克隆抗体制备

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摘要

Differentially expressed genes between normal SPG-A1 cells and SPC-A1 cells infected by RSV were investigated using differential display. The novel z1g10 gene codes for a novel protein, ZLG10, which has previously been reported to be up-regulated in RSV-infected SPC-A1 cells. Its putative open reading frame was also identified. To better understand the structure, function, and possible role of ZLG10 as a potential candidate for diagnosis and vaccine studies, the intact region encoding ZLG10 was obtained by PCR and expressed in Escherichia coli as a GST-fusion protein. After purification, GST-ZLG10 fusion protein was used to immunize the adult rabbits following standard protocols. Consequently, we found that the produced antiserum of the novel fusion protein significantly suppressed the infection by RSV on SPC-A1 cells by using neutral red uptake assay and quantitative measurement. Together, our data demonstrate that ZLG10, a novel protein expressed and purified in this report, might be a potential effective therapeutic candidate for treating RSV infections. (c) 2005 Elsevier Inc. All rights reserved.
机译:使用差异显示技术研究了正常SPG-A1细胞和受RSV感染的SPC-A1细胞之间差异表达的基因。新的z1g10基因编码一种新蛋白ZLG10,以前据报道该蛋白在RSV感染的SPC-A1细胞中被上调。还确定了其推定的开放阅读框。为了更好地了解ZLG10作为诊断和疫苗研究的潜在候选者的结构,功能和可能的作用,通过PCR获得了编码ZLG10的完整区域,并在大肠杆菌中以GST融合蛋白的形式表达。纯化后,使用GST-ZLG10融合蛋白按照标准规程免疫成年兔。因此,我们发现所产生的新型融合蛋白的抗血清通过使用中性红吸收测定和定量测量,显着抑制了RSV对SPC-A1细胞的感染。总之,我们的数据表明ZLG10是在本报告中表达和纯化的一种新型蛋白质,可能是治疗RSV感染的潜在有效治疗候选物。 (c)2005 Elsevier Inc.保留所有权利。

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