Cxcl16 与 SARS-CoV N 蛋白在细胞内外相互作用。
Cxcl16 interact with SARS-CoV N protein in and out cell.
机构信息
College of Animal Science and Technology, Shandong Agricultural University, Tai'an, 271018, China.
出版信息
Virol Sin. 2010 Oct;25(5):369-74. doi: 10.1007/s12250-010-3129-x. Epub 2010 Oct 8.
Abstract
Our study investigated the host cell protein which can interact with SARS-CoV N protein, and explored the functional connections. The eukaryotic expression vectors pEGFP-N1/SARS-CoVN and pdsRed2-N1/CXCL16 were constructed and used to co-transfect HEK293FT cells by the calcium phosphate method. The HIS-tagged fusion protein SARS-CoVN-GFP was then built and purified for the binding assay in vitro. The co-localization of SARS-CoVN and CXCL16 in the cytoplasm of HEK293FT cells was also shown using confocal laser scanning microscopy. It is suggested that their interaction might be through direct combination. Under a fluorescence microscope, it was observed that the purified fusion protein SARS-CoVN-GFP was attached to the cell membrane of CXCL16-transfected cells, indicating that SARS-CoVN and CXCL16 can be mutually combined.
摘要
我们的研究调查了可以与 SARS-CoV N 蛋白相互作用的宿主细胞蛋白，并探讨了它们的功能联系。构建了真核表达载体 pEGFP-N1/SARS-CoVN 和 pdsRed2-N1/CXCL16，并用磷酸钙法共转染 HEK293FT 细胞。然后构建了带有 HIS 标签的融合蛋白 SARS-CoVN-GFP，并进行体外结合实验进行纯化。利用共聚焦激光扫描显微镜观察到 SARS-CoVN 和 CXCL16 在 HEK293FT 细胞质中的共定位。提示它们的相互作用可能是通过直接结合。在荧光显微镜下，观察到纯化的融合蛋白 SARS-CoVN-GFP 附着在 CXCL16 转染细胞的细胞膜上，表明 SARS-CoVN 和 CXCL16 可以相互结合。
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