In Vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate

doi:10.1007/978-1-62703-782-2_13

科研成果详情

题名	In Vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate
作者	Zhao, Kong-Nan1,2
发表日期	2014
发表期刊	CELL-FREE PROTEIN SYNTHESIS: METHODS AND PROTOCOLS 影响因子和分区
语种	英语
原始文献类型	Article ; Book Chapter
关键词	Primary mouse keratinocytes Cell-free lysate Codon usage Aminoacyl-tRNAs Papillomavirus L1 gene mRNAs In vitro translation
其他关键词	SKIN BARRIER FUNCTION ; PROINFLAMMATORY MEDIATORS ; PROTEIN EXPRESSION ; FREE SYSTEM ; DIFFERENTIATION ; SEQUENCES ; USAGE ; DNA ; TRANSCRIPTION ; FILAGGRIN
摘要	Keratinocytes are the major cell type in the epidermis responsible for constructing the protective barrier of mammalian skin. Primary keratinocytes cultured in vitro can mimic the in vivo differentiation process, providing an abundant source of pure keratinocytes used for investigating the regulatory mechanisms of gene expression associated with cell proliferation and differentiation. We developed a primary mouse keratinocyte cell-free lysate system for translation of papillomavirus authentic and codon-modified L1 capsid gene mRNAs in vitro. We demonstrated that the viral gene codon usage matched available aminoacyl-tRNAs to determine their translational efficiencies, which were associated with differentiation status of the keratinocytes used for preparing cell-free lysate. We revealed that a novel regulatory mechanism of gene expression is utilized by papillomavirus to direct viral capsid protein expression to the site of virion assembly in mature keratinocytes. Here, I describe the methods in details of how to establish primary mouse keratinocyte culture, to prepare cell-free lysate, to carry out in vitro translation of the viral gene mRNAs, and to detect the translated products using Western blotting analysis.
资助项目	National Health and Medical Research Council[undefined];
出版者	HUMANA PRESS INC
出版地	TOTOWA
ISSN	1064-3745
EISSN	1940-6029
卷号	1118 页码:205-218
DOI	10.1007/978-1-62703-782-2_13
页数	14
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology
WOS研究方向	Biochemistry & Molecular Biology
WOS记录号	WOS:000329751700014
收录类别	BKCI-S ; SCOPUS
URL	查看原文
PubMed ID	24395418
SCOPUSEID	2-s2.0-84934443010
通讯作者地址	[Zhao, Kong-Nan]Institute of Molecular Virology and Immunology, Wenzhou Medical College,China
Scopus学科分类	Molecular Biology;Genetics
引用统计	被引频次：1[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/824
专题	检验医学院（生命科学学院、生物学实验教学中心）_病原生物学与免疫学系
通讯作者	Zhao, Kong-Nan
作者单位	1.UQ Centre for Clinical Research, University of Queensland,Australia; 2.Institute of Molecular Virology and Immunology, Wenzhou Medical College,China
第一作者单位	温州医科大学
通讯作者单位	温州医科大学
推荐引用方式 GB/T 7714	Zhao, Kong-Nan. In Vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate[J]. CELL-FREE PROTEIN SYNTHESIS: METHODS AND PROTOCOLS,2014,1118:205-218.
APA	Zhao, Kong-Nan. (2014). In Vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate. CELL-FREE PROTEIN SYNTHESIS: METHODS AND PROTOCOLS, 1118, 205-218.
MLA	Zhao, Kong-Nan."In Vitro Translation of Papillomavirus Authentic and Codon-Modified L1 Capsid Gene mRNAs in Mouse Keratinocyte Cell-Free Lysate".CELL-FREE PROTEIN SYNTHESIS: METHODS AND PROTOCOLS 1118(2014):205-218.