20210317

Contents
  1. 1. Working directories
    1. 1.1. CEB format
      1. 1.1.1. CEB 文件摘要
      2. 1.1.2. CEB/Word conversion
    2. 1.2. Script in ht_5T_9 and ht_T7_1
  2. 2. Hexo tips
  3. 3. Virus proliferation cell free system
    1. 3.1. Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube
      1. 3.1.1. Abstract
    2. 3.2. Cell-free RNA replication systems based on a human cell extracts-derived in vitro translation system with the encephalomyocarditisvirus RNA
      1. 3.2.1. Abstract
    3. 3.3. An improved cell-free system for picornavirus synthesis
      1. 3.3.0.1. Erratum in
    4. 3.3.1. Abstract
  4. 3.4. Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube
    1. 3.4.1. Abstract
  5. 3.5. A Cell-Free Assembly System for Generating Infectious Human Papillomavirus 16 Capsids Implicates a Size Discrimination Mechanism for Preferential Viral Genome Packaging
    1. 3.5.1. ABSTRACT
  6. 3.6. Cell-free synthesis of encephalomyocarditis virus
    1. 3.6.0.1. Abstract

Working directories

CEB format

CEB 文件摘要

根据我们的记录，有 一 种与 CEB 文件扩展名相关的文件类型，最常见的被格式化为 Apabi eBook File。Apabi Reader 由 Founder Group 发布，是最通用的关联应用程序。 此外，有 一 种不同的软件程序可供您用于查看这些文件。 大多数 CEB 文件被视为 eBook Files。

CEB 文件可以在移动和桌面平台上找到，可以在 Windows中打开。 这些文件的普及性为“低”，这意味着它们并不常见。

如果您在打开 CEB 文件时遇到问题，或者只是想了解有关它们的软件程序和开发人员的更多信息，请参阅下面的完整信息。

CEB/Word conversion

Script in ht_5T_9 and ht_T7_1

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i='/media/ht/ht_5T_9/YouTubes/README.sh'; subl ${i}
i='/media/ht/ht_T7_1/README.sh'; subl ${i}
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# y2mate.com
#
space; space; rename 's/\_720p//; s/\_240p//; s/\_360p//; s/\_480p//; s/\_1080p//; s/\_v720P//'  y2*0p.* y2*0P.*
Remove_cnmark; Remove_cnmark
#
for i in y2m*.*; do mv ${i} ${i:11:200}; done
for i in y2m*.*; do mv ${i} ${i:11:200}; done; for i in *.*; do mv ${i} `echo ${i} | rev | cut -b 17-100 | rev`.${i#*.*}; done

Hexo tips

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i='https://segmentfault.com/a/1190000017986794 https://hexo.io/zh-cn/docs/github-pages.html https://www.jianshu.com/p/7daa1dc0e117'; setsid chromium-browser ${i}

Virus proliferation cell free system

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i='https://jvi.asm.org/content/90/17/7607/article-info https://jvi.asm.org/content/jvi/90/17/7607.full.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3838232/ https://pubmed.ncbi.nlm.nih.gov/21952914/ https://pubmed.ncbi.nlm.nih.gov/25488519/ https://pubmed.ncbi.nlm.nih.gov/21622667/ https://pubmed.ncbi.nlm.nih.gov/17320977/ https://pubmed.ncbi.nlm.nih.gov/12743313/ https://link.springer.com/article/10.1007%2Fs10529-011-0744-z'; setsid chromium-browser ${i}

Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube

Tominari Kobayashi 1, Yusuke Nakamura, Satoshi Mikami, Mamiko Masutani, Kodai Machida, Hiroaki Imataka

Affiliations expand

Abstract

Virus particles are used in vaccination, drug delivery, and material sciences. Here we devised a system where the RNA virus encephalomyocarditis virus (EMCV) is synthesized from DNA templates in vitro. When a plasmid or a PCR product harboring the full-length cDNA of EMCV in the T7 promoter/terminator unit was incubated in a HeLa cell extract supplemented with T7 RNA polymerase, EMCV was produced within 4 h at an efficiency of over 10-fold compared with the system programmed with the EMCV RNA. The EMCV RNA transcribed by the virally encoded RNA-dependent RNA polymerase was predominantly incorporated into the EMCV particle even in the presence of a larger amount of the EMCV RNA transcribed by T7 RNA polymerase from the plasmid.

Cell-free RNA replication systems based on a human cell extracts-derived in vitro translation system with the encephalomyocarditisvirus RNA

Tominari Kobayashi 1, Kodai Machida, Satoshi Mikami, Mamiko Masutani, Hiroaki Imataka

Affiliations expand

Abstract

To study the relationship between translation and replication of encephalomyocarditisvirus (EMCV) RNA, we established a cell-free RNA replication system by employing a human cell extracts-based in vitro translation system. In this system, a cis-EMCV RNA replicon encoding the Renilla luciferase (R-luc) or GFP and the viral regulatory proteins efficiently replicated with simultaneous translation of the encoded protein. To examine how translation of the replicon RNA, but not the translated products, affected replication, a trans-EMCV RNA replicon encoding R-luc and the RNA replication elements was next constructed. The trans-replicon RNA replicated only in the presence of the regulatory proteins pre-expressed in trans. Incubation with cycloheximide, puromycin or a dominant-negative eukaryotic translation initiation factor 4A following expression of the regulatory proteins almost completely inhibited not only translation of the trans-replicon RNA but also replication of the RNA, suggesting that EMCV RNA translation promotes replication of the RNA. In conclusion, the cell-free RNA replication systems should become useful tools for the study of the viral RNA replication.

An improved cell-free system for picornavirus synthesis

Tominari Kobayashi 1, Satoshi Mikami, Shigeyuki Yokoyama, Hiroaki Imataka

Affiliations expand

Erratum in

  • J Virol Methods. 2012 Jan;179(1):e1

Abstract

Cell-free synthesis of an infectious virus is an ideal tool for elucidating the mechanism of viral replication and for screening anti-viral drugs. In the present study, the synthesis of Encephalomyocarditis virus (EMCV) from its RNA in HeLa and 293-F cell extracts was enhanced by employing a dialysis system in combination with a ribozyme technology. Although translation and processing of the EMCV polyprotein were not accelerated greatly by the dialysis system, de novo synthesis of viral RNA was enhanced considerably by dialysis, leading to a greater than eight-fold increased titer of synthesized EMCV compared with a conventional batch system. Furthermore, a synthetic EMCV RNA with a hammerhead ribozyme sequence at its 5’-end served as an efficient template for viral synthesis in the dialysis system. Therefore, this system provides opportunities for mutational analyses of EMCV in vitro.

Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube

Biotechnology Letters volume 34, pages67–73(2012)Cite this article

Abstract

Virus particles are used in vaccination, drug delivery, and material sciences. Here we devised a system where the RNA virus encephalomyocarditis virus (EMCV) is synthesized from DNA templates in vitro. When a plasmid or a PCR product harboring the full-length cDNA of EMCV in the T7 promoter/terminator unit was incubated in a HeLa cell extract supplemented with T7 RNA polymerase, EMCV was produced within 4 h at an efficiency of over 10-fold compared with the system programmed with the EMCV RNA. The EMCV RNA transcribed by the virally encoded RNA-dependent RNA polymerase was predominantly incorporated into the EMCV particle even in the presence of a larger amount of the EMCV RNA transcribed by T7 RNA polymerase from the plasmid.

A Cell-Free Assembly System for Generating Infectious Human Papillomavirus 16 Capsids Implicates a Size Discrimination Mechanism for Preferential Viral Genome Packaging

Carla Cerqueira, Yuk-Ying S. Pang, Patricia M. Day, Cynthia D. Thompson, Christopher B. Buck, Douglas R. Lowy, John T. Schiller

S. Perlman, Editor

DOI: 10.1128/JVI.02497-15

ABSTRACT

We have established a cell-free in vitro system to study human papillomavirus type 16 (HPV16) assembly, a poorly understood process. L1/L2 capsomers, obtained from the disassembly of virus-like particles (VLPs), were incubated with nuclear extracts to provide access to the range of cellular proteins that would be available during assembly within the host cell. Incorporation of a reporter plasmid “pseudogenome” was dependent on the presence of both nuclear extract and ATP. Unexpectedly, L1/L2 VLPs that were not disassembled prior to incubation with a reassembly mixture containing nuclear extract also encapsidated a reporter plasmid. As with HPV pseudoviruses (PsV) generated intracellularly, infection by cell-free particles assembled in vitro required the presence of L2 and was susceptible to the same biochemical inhibitors, implying the cell-free assembled particles use the infectious pathway previously described for HPV16 produced in cell culture. Using biochemical and electron microscopy analyses, we observed that, in the presence of nuclear extract, intact VLPs partially disassemble, providing a mechanistic explanation to how the exogenous plasmid was packaged by these particles. Further, we provide evidence that capsids containing an <8-kb pseudogenome are resistant to the disassembly/reassembly reaction. Our results suggest a novel size discrimination mechanism for papillomavirus genome packaging in which particles undergo iterative rounds of disassembly/reassembly, seemingly sampling DNA until a suitably sized DNA is encountered, resulting in the formation of a stable virion structure.

IMPORTANCE Little is known about papillomavirus assembly biology due to the difficulties in propagating virus in vitro. The cell-free assembly method established in this paper reveals a new mechanism for viral genome packaging and will provide a tractable system for further dissecting papillomavirus assembly. The knowledge gained will increase our understanding of virus-host interactions, help to identify new targets for antiviral therapy, and allow for the development of new gene delivery systems based on _in vitro_-generated papillomavirus vectors.

Cell-free synthesis of encephalomyocarditis virus

Yuri V Svitkin 1, Nahum Sonenberg

Affiliations expand

Free PMC article

Abstract

We developed a system for complete replication of encephalomyocarditis virus (EMCV) in a test tube by using an in vitro translation extract from Krebs-2 cells. Efficient virus synthesis occurred in a narrow range of Mg(2+) and EMCV RNA concentrations. Excess input RNA impaired RNA replication and virus production but not translation. This suggests the existence of a negative-feedback mechanism for regulation of RNA replication by the viral plus-strand RNA or proteins.

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    缺失模块。
    1、请确保node版本大于6.2
    2、在博客根目录（注意不是yilia根目录）执行以下命令：
    npm i hexo-generator-json-content --save

    3、在根目录_config.yml里添加配置： 

      jsonContent:
    meta: false
    pages: false
    posts:
      title: true
      date: true
      path: true
      text: false
      raw: false
      content: false
      slug: false
      updated: false
      comments: false
      link: false
      permalink: false
      excerpt: false
      categories: false
      tags: true

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