GB病毒C基因7型重组E2蛋白的克隆、序列分析及表达

Xiaoyu Yang, Yue Zhao, Yue Feng, Xueshan Xia
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引用次数: 0

摘要

本研究的目的是探讨GB病毒C(GBV-C)基因型7E2蛋白作为ELISA试剂盒检测抗原的潜力。本研究使用在线分析程序对GBV-C基因型7E2蛋白的抗原表位、空间结构和线性B细胞表位进行了分析。为了建立更可靠的GBV-C研究检测方法,采用RT-PCR扩增出GBV-C E2基因945bp片段,并连接到原核表达载体pET-32中,转化大肠杆菌BL21细胞进行蛋白表达。12% SDS-PAGE检测到分子量为55 kDa的蛋白。在包涵体中发现了该蛋白,并用western blotting检测了his标记蛋白。结果表明,克隆的E2基因序列为945 bp,且GBV-C E2蛋白序列具有多个抗原表位。重组蛋白形成包涵体,与预期一致。这些发现可能为GBV-C检测试剂盒的开发提供基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Cloning, Sequence Analysis and Expression of Recombinant E2 Protein of GB Virus C Genotype 7].

The purpose of this study was to explore the potential of the GB virus C(GBV-C)genotype 7E2 protein as a detection antigen for ELISA kit development. In this study, analyses of antigen epitopes, space structures and the linear B cell epitopes from the GBV-C genotype 7E2 protein were performed using an online analysis program. To establish a more reliable detection method for GBV-C studies, a 945bp gene fragment from GBV-C E2 was amplified by RT-PCR and ligated into the pET-32 a prokaryotic expression vector, which was then transformed into E. coli BL21 cells for protein expression. A protein with a molecular weight of 55 kDa was detected by 12% SDS-PAGE. The protein was found in inclusion bodies, and the His-tagged protein was detected by western blotting. The results showed that the cloned E2 gene sequence was 945 bp, and that the GBV-C E2 protein sequence had multiple antigenic epitopes. The recombinant protein formed inclusion bodies, which was consistent with expectations. These findings may provide the foundation for the development of a GBV-C detection kit.

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