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当前位置: 首页 > 文章 > CBL蛋白的真核表达及其对日本脑炎病毒的抑制作用 畜牧与兽医 2018 (1) 86-90
Position: Home > Articles > Eukaryotic expression of CBL protein and its inhibition against Japanese encephalitis virus Animal Husbandry & Veterinary Medicine 2018 (1) 86-90

CBL蛋白的真核表达及其对日本脑炎病毒的抑制作用

作  者：
梁静泊;高帅;申坤;李本睿;宗嫚嫚;郑阳;余远楠;谢金峰;冯秀丽
单  位：
南京农业大学动物医学院
关键词：
CBL;真核表达;日本脑炎病毒(JEV);抗病毒作用
摘  要：
CBL蛋白是一种E3泛素连接酶,在调控信号传导等方面发挥重要作用。采用PCR扩增CBL基因片段并连接到pEGFP-C1载体中,经Bam HⅠ、HindⅢ双酶切鉴定及基因测序,成功构建真核表达重组载体pEGFP-CBL。倒置荧光显微镜观察和Western blot结果证实,重组载体pEGFP-CBL在BHK-21细胞中成功表达。病毒增殖试验结果证实,日本脑炎病毒(JEV)强毒株感染转染pEGFP-CBL重组载体的BHK-21细胞,3 d内未见明显细胞病变。结果表明,真核表达载体pEGFP-CBL成功构建,且重组表达的CBL蛋白能够抑制早期JEV增殖,从而为深入研究CBL蛋白的免疫调节功能奠定了基础。
译  名：
Eukaryotic expression of CBL protein and its inhibition against Japanese encephalitis virus
作  者：
LIANG Jingbo;GAO Shuai;SHEN Kun;LI Benrui;ZONG Manman;ZHENG Yang;YU Yuannan;XIE Jinfeng;FENG Xiuli;College of Veterinary Medicine,Nanjing Agriculture University;
关键词：
CBL;;eukaryotic expression;;Japanese encephalitis virus(JEV);;antiviral proliferation
摘  要：
Casitas B-lineage lymphoma(CBL) is a E3 ubiquitin ligase,which is known to play an important role in regulating signal transduction,receptor internalization and signal termination.In this paper,CBL fragment was amplified and introduced into the pEGFP-C1 vector.After being confirmed by Bam HⅠ and Hind Ⅲ digestion analyses and DNA sequencing,the recombinant pEGFP-CBL plasmid was transfected into BHK-21 cells.The expression of the recombinant plasmid pEGFP-CBL in BHK21 cells and its antiviral activity was detected by fluorescence microscopy,western blotting and inhibition of viral proliferation assay,respectively.Results showed the recombinant pEGFP-CBL was effectively expressed in BHK-21 cells.BHK-21 cells transfected with pEGFP-CBL recombinant vector did not show significant cytopathic effect within 3 days after infection with Japanese encephalitis virus(JEV),which suggested that a recombinant vector pEGFP-CBL was successfully constructed and expressed,and CBL protein might inhibit early JEV proliferation.This study laid the foundation for the further investigation on the immunomodulatory function of CBL protein.

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