荧光蛋白标记的乙型肝炎病毒衣壳蛋白与支持核衣壳形成的

StephenW

J Virol Methods. 2018 Feb 12. pii: S0166-0934(17)30556-6. doi: 10.1016/j.jviromet.2018.02.010. [Epub ahead of print]
Fluorescent protein tagged hepatitis B virus capsid protein with long glycine-serine linker that supports nucleocapsid formation.Chen JY1, Gan CY2, Cai XF2, Zhang WL2, Long QX2, Wei XF2, Hu Y2, Tang N2, Chen J2, Guo H3, Huang AL4, Hu JL5.
Author information
1Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China; Department of Clinical Laboratory, The First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (CCID), Hangzhou, China.2Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.3Department of Microbiology and Immunology, Indiana University School of Medicine, United States.4Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (CCID), Hangzhou, China. Electronic address: [email protected].5Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases (CCID), Hangzhou, China. Electronic address: [email protected].

AbstractFusion core proteins of Hepatitis B virus can be used to study core protein functions or capsid trafficking. A problem in constructing fusion core proteins is functional impairment of the individual domains in these fusion proteins, might due to structural interference. We reported a method to construct fusion proteins of Hepatitis B virus core protein (HBc) in which the functions of fused domains were partially kept. This method follows two principles: (1) fuse heterogeneous proteins at the N terminus of HBc; (2) use long Glysine-serine linkers between the two domains. Using EGFP and RFP as examples, we showed that long flexible G4S linkers can effectively separate the two domains in function. Among these fusion proteins constructed, GFP-G4S186-HBc and RFP-G4S47-HBc showed the best efficiency in rescuing the replication of an HBV replicon deficient in the core protein expression, though both of the two fusion proteins failed to support the formation of the relaxed circular DNA. These fluorescent protein-tagged HBcs might help study related to HBc or capsids tracking in cells.


KEYWORDS: Hepatitis B virus; core protein; engineering; fusion proteins

PMID:29447911DOI:10.1016/j.jviromet.2018.02.010

StephenW

Virol方法。 2018年2月12日。pii：S0166-0934（17）30556-6。 doi：10.1016 / j.jromerome.2018.02.010。 [电子版提前打印]
荧光蛋白标记的乙型肝炎病毒衣壳蛋白与支持核衣壳形成的长甘氨酸 - 丝氨酸接头。
Chen JY1，Gan CY2，Cai XF2，Zhang WL2，Long QX2，Wei XF2，Hu Y2，Tang N2，Chen J2，Guo H3，Huang AL4，Hu JL5。
作者信息

1
    重庆医科大学附属第二医院感染科传染病分子生物学教育部重点实验室;汕头大学医学院附属第一医院检验科，广东汕头;中国杭州传染病诊疗协作创新中心（CCID）。
2
    重庆医科大学附属第二医院感染科传染病分子生物学教育部重点实验室，重庆，中国。
3
    美国印第安纳大学医学院微生物学与免疫学系。
4
    重庆医科大学附属第二医院感染科传染病分子生物学教育部重点实验室;中国杭州传染病诊疗协作创新中心（CCID）。电子地址：[email protected]。
五
    重庆医科大学附属第二医院感染科传染病分子生物学教育部重点实验室;中国杭州传染病诊疗协作创新中心（CCID）。电子地址：[email protected]。

抽象

乙型肝炎病毒融合核心蛋白可用于研究核心蛋白功能或衣壳运输。构建融合核心蛋白质的问题是这些融合蛋白中单个结构域的功能损伤，可能是由于结构干扰。我们报道了构建乙型肝炎病毒核心蛋白（HBc）融合蛋白的方法，其中融合结构域的功能部分保留。该方法遵循两个原则：（1）在HBc的N端融合异源蛋白质; （2）在两个结构域之间使用长的甘氨酸 - 丝氨酸连接子。以EGFP和RFP为例，我们发现长期灵活的G4S连接子可以有效地分离两个功能域。在所构建的这些融合蛋白中，GFP-G4S186-HBc和RFP-G4S47-HBc在拯救HBV复制子核心蛋白表达缺陷的复制方面显示出最佳效率，尽管两种融合蛋白都不能支持形成放松的环状DNA。这些荧光蛋白标记的HBcs可能有助于研究HBc或细胞中的衣壳跟踪。
关键词：

乙型肝炎病毒;核心蛋白;工程;融合蛋白

结论：
    29447911
DOI：
    10.1016 / j.jviromet.2018.02.010

StephenW

https://www.sciencedirect.com/sc ... i/S0166093417305566