In vitro translation of papillomavirus authentic and codon-modified L1 capsid gene mRNAs in mouse keratinocyte cell-free lysate

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-Nan
著作类型	图书著作
出版日期	2014
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. 2014 Springer Science+Business Media, LLC.
ISBN	97816270
语种	英语
出版单位	Methods in Molecular Biology
文献类型	专著
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/101284
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
作者单位	1.UQ Centre for Clinical Research, University of Queensland, Herston, Brisbane, QLD, Australia 2.Institute of Molecular Virology and Immunology, Wenzhou Medical College, Wenzhou, Zhejiang, China
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[M],2014.