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Rapid, Efficient, and Modular Generation of Adenoviral Vectors via Isothermal Assembly.
Yang, Yong; Chi, Yudan; Tang, Xinying; Ertl, Hildegund C J; Zhou, Dongming.
Afiliação
  • Yang Y; Vaccine Research Center of Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
  • Chi Y; Vaccine Research Center of Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
  • Tang X; Vaccine Research Center of Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
  • Ertl HCJ; The Wistar Institute, Philadelphia, Pennsylvania.
  • Zhou D; Vaccine Research Center of Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
Curr Protoc Mol Biol ; 113(1): 16.26.1-16.26.18, 2016 Jan.
Article em En | MEDLINE | ID: mdl-31773916
ABSTRACT
Adenoviral vectors have yielded promising results as carriers for gene transfer and vaccines in basic research and clinical applications. However, most common procedures to construct adenoviral vectors and manipulate adenovirus (Ad) genomes are complex and labor-intensive. An easy and detailed protocol for the rapid, efficient, and modular generation of chimpanzee Ad serotype 68 (AdC68) as a molecular clone via isothermal assembly, which directionally assembles multiple DNA fragments in a single isothermal reaction without restriction enzymes or ligases, is presented. Any serotype of adenovirus with the sequence of genome known can be constructed as a molecular clone by this method. Recombinant adenoviral vectors can be created via one-step isothermal assembly in <3 days, and recombinant Ads can be rescued within 8 days. This protocol is practical for manipulations of Ad genomes, because an entire Ad genome can be divided into specific fragments within modular plasmids. © 2016 by John Wiley & Sons, Inc.
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Texto completo: 1 Base de dados: MEDLINE Tipo de estudo: Guideline Idioma: En Ano de publicação: 2016 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Tipo de estudo: Guideline Idioma: En Ano de publicação: 2016 Tipo de documento: Article