Nonstructural proteins of Torque teno sus virus 2 from O2AUG: Prediction to experimental validation
作者： Yan Lu , Haisheng Yu , Xiaohua Nie , Yongqing Li , Liguo Zhang
DOI: 10.1016/J.VIRUSRES.2013.09.031
关键词:
摘要: The expression profiles of nonstructural proteins (NSPs) in Torque teno sus virus 2 (TTSuV2) have not yet been characterized. Here, we determined the coding sequences TTSuV2 NSPs ORF2, ORF2/2, and ORF2/2/3 by overlapping polymerase chain reaction (PCR) subsequent bacterial mammalian cells. We generated two monoclonal antibodies (mAbs), 2E5 6F8, from mice immunized with mixed Escherichia coli expressing His-tagged ORF2 ORF2/2. Enzyme-linked immunosorbent assay (ELISA) western blot analysis revealed that, mAbs bound to consensus ORF2/2/3, while 6F8 recognized common ORF2/2 ORF2/2/3. Immunofluorescence (IFA) that was localized cytoplasm, nucleus but nucleolus, peri-nuclear region. To identify TTSuV NSPs, a circular TTSuV2_ZJ (GenBank: KF660540) genomic DNA clone constructed transfected into HEK293T HeLa Splicing mRNAs localization were identified RT-PCR, analysis, IFA, respectively. However, detected either at RNA or protein level. Our study is first provide experimental evidence existence Moreover, potential applications future research on viral function diagnosis related diseases.
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