膜联蛋白A2对脑心肌炎病毒在BHK-21细胞中增殖的影响

发布时间：2018-11-26 20:21

【摘要】：膜联蛋白A2是一种参与调节多种病毒增殖的重要宿主蛋白。本研究通过构建过表达膜联蛋白A2的BHKAnxa2细胞系及瞬时敲低膜联蛋白A2,探讨膜联蛋白A2对脑心肌炎病毒在BHK-21细胞中增殖的影响。通过RT-PCR从BHK-21细胞中扩增膜联蛋白A2全长cDNA,测序正确后克隆入pcDNA3.1整合表达载体中;用重组载体pcDNA3.1-Anxa2转染BHK-21细胞,经G418筛选获得抗性克隆,qRT-PCR和Western blot试验证明BHKAnxa2过表达膜联蛋白A2;EMCV感染试验证明BHK-Anxa2细胞中病毒滴度高于对照组。通过siRNA降低BHK-21细胞中膜联蛋白A2表达,EMCV感染试验证明膜联蛋白A2表达下降后EMCV增殖也随之下降。以上试验结果表明鼠膜联蛋白A2表达改变可导致病毒在BHK-21细胞中的增殖发生变化,提示膜联蛋白A2与EMCV在BHK-21细胞中的增殖相关。
[Abstract]:Annexin A2 is an important host protein involved in regulating the proliferation of many viruses. The aim of this study was to investigate the effect of BHKAnxa2 cell line and transient knockout A2 on the proliferation of encephalitis virus in BHK-21 cells. The full length cDNA, of annexin A2 was amplified by RT-PCR from BHK-21 cells and cloned into pcDNA3.1 integrative expression vector. The recombinant vector pcDNA3.1-Anxa2 was used to transfect BHK-21 cells, and the resistant clones were obtained by G418 screening. QRT-PCR and Western blot tests showed that the viral titer of BHK-Anxa2 cells was higher than that of the control group. The results of qRT-PCR and Western blot showed that the viral titer of BHK-Anxa2 cells was higher than that of the control group. The expression of annexin A2 in BHK-21 cells was decreased by siRNA, and the EMCV proliferation was also decreased after the decrease of EMCV expression by EMCV infection assay. These results suggest that the expression of rat integrin A2 may lead to the proliferation of the virus in BHK-21 cells, suggesting that there is a correlation between the proliferation of EMCV and the proliferation of integrin A2 in BHK-21 cells.
【作者单位】：甘肃省动物细胞工程技术研究中心;西北民族大学生命科学与工程学院;西北民族大学生物工程与技术国家民委重点实验室;
【基金】：国家自然科学基金(项目号:31460665),题目:脑心肌炎病毒受体的筛选、鉴定及功能分析教育部“长江学者和创新团队发展计划”(项目号:IRT13091) 中央高校基本科研业务费专项资金(项目号:31920150025),题目:膜联蛋白 A2调节脑心肌炎病毒感染的作用和机制研究。
【分类号】：R373

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