摘要
为研究马立克病病毒（Marek＇s disease virus,MDV）致瘤蛋白Meq对细胞周期蛋白依赖性激酶抑制蛋白p21表达的调节作用,采用RTPCR、Western blot方法检测Meq蛋白表达在鸡胚成纤维细胞（Chicken embryo fibroblasts,CEF）中对p21 mRNA和蛋白水平的影响。然后,构建p21启动子的荧光素酶报告载体,检测Meq蛋白对p21的启动子活性的影响。最后,分别用MDV Md5强毒株和CVI988/Rispens疫苗毒株感染CEF细胞,利用Western blot检测MDV感染后p21蛋白水平的变化。结果：过表达Meq上调p21的mRNA和蛋白表达水平,Meq能够激活p21基因启动子活性,MDV感染细胞中p21蛋白的表达水平也有明显升高。这些发现为了解Meq在MDV感染中的作用机理奠定了基础。
To investigate whether Marek＇s disease virus （MDV） Meq oncoprotein could regulate cyclin dependent kinase inhibitor p21, RT-PCR and Western Blotting were first performed to examine the regulation of mRNA and protein levels of p21 by Meq overexpression. Then, the p21 promoter was analyzed and cloned into the luciferase reporter vector, and fold change of luciferase activity of the p21 promoter upon Meq overexpression was examined. Finally, p21 protein levels upon infection of MDV Md5 and CVI988/Rispens strains were checked. The results showed that p21 mRNA and protein levels were upregulated by Meq, that Meq overexpression activated the p21 promoter, and that MDV infection increased p21 protein levels. These findings together lay an important foundation for understanding the role of Meq in MDV infection.
作者
鲍晨沂
连雪
杨莎莎
张旭
JUNG Yong-Sam
钱莺娟
BAO Chenyi;LIAN Xue;YANG Shasha;ZHANG Xu;JUNG Yong-Sam;QIAN Yingjuan(Key Lab of Animal Bacteriology, Ministry of Agriculture/MOE Joint International Research Laboratory of Animal Health and Food Safety/College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China)
出处
《畜牧与兽医》
北大核心
2018年第5期84-88,共5页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31472218)
江苏省自然科学青年基金(BK20140711)
江苏省优势学科专项经费
