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作 者:刘俊丽[1] 冯怡[1] 李楚芳[1] 曹诚[1] 马清钧[1]
机构地区:[1]军事医学科学院生物工程研究所,北京100850
出 处:《生物技术通讯》2006年第2期138-141,共4页Letters in Biotechnology
基 金:国家高技术研究发展计划重大专项(2005AA218080)
摘 要:目的:SARS冠状病毒(SARS-CoV)的核衣壳蛋白(N蛋白)能够结合病毒RNA形成螺旋状的核衣壳。在3种不同的细胞系中分别表达SARS-CoVN蛋白,研究它对转染细胞生长的影响。方法:将重组质粒pEGFP-N和pEGFP(作为对照)分别转染人胚肾HEK293细胞、成纤维细胞3T3、人宫颈癌HeLa细胞,通过激光共聚焦显微镜、荧光显微镜观察SARS-CoVN蛋白在细胞内的定位以及细胞的生长变化。结果:SARS-CoVN蛋白能定位于细胞质,并不像其他冠状病毒N蛋白那样能够定位到细胞核。同时发现SARS-CoVN蛋白能够诱导形成多核细胞,多核细胞的百分率可达33.9%。结论:SARS-CoVN蛋白抑制胞质分裂,延缓细胞生长。Objective: SARS coronavirus(SARS-CoV) Nucleocapsid (N) proein is a structural protein of SARS-CoV, which binds viral RNA to form the helical nucleocapsid. In this study, SARS-CoV N protein was respectively expressed in three different cell lines in order to investigate its influence on the growth of cells expressing the N protein of SARS- CoV. Methods: Plasmid pEGFP-N and pEGFP (as control) were transfected respectively into HEK293 cells, 3T3 cells and HeLa cells. The localization of SARS-CoV N protein and the change of its growth were examined by fluorescent microscopy and confocal microscopy. Results: It was found that the SARS-CoV N protein was localized in cytoplasma other than nucleolear, which was found in the N protein of other coronaviruses. More over, it was found that N protein of SARS-CoV induced multinucleated cells. The percentage of multinucleated cells was as high as 33.9%. Conclusion: Cytokinesis was inhibited and cell growth was delayed by the N protein of the SARS-CoV.
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