Generation of recombinant coronaviruses using vaccinia virus as the cloning vector and stable cell lines containing coronaviral replicon RNAs

Publikation

Generation of recombinant coronaviruses using vaccinia virus as the cloning vector and stable cell lines containing coronaviral replicon RNAs

Buchkapitel - 01.01.2008

Eriksson Klara Kristin, Makia Divine, Thiel Volker

Zitation

Eriksson K, Makia D, Thiel V. Generation of recombinant coronaviruses using vaccinia virus as the cloning vector and stable cell lines containing coronaviral replicon RNAs. In: SARS and Other Coronaviruses. Totowa, New Jersey: Humana Press, 2008. S. 237-54.

Art

Buchkapitel (Englisch)

Buchtitel

SARS and Other Coronaviruses

Veröffentlichungsdatum

01.01.2008

Titel der Reihe

Methods in Molecular Biology (454)

Seiten

237-54

Verlag

Humana Press (Totowa, New Jersey)

Kurzbeschreibung/Zielsetzung

Coronavirus reverse genetic systems have become valuable tools for studying the molecular biology of coronavirus infections. They have been applied to the generation of recombinant coronaviruses, selectable replicon RNAs, and coronavirus-based vectors for heterologous gene expression. Here we provide a collection of protocols for the generation, cloning, and modification of full-length coronavirus cDNA using vaccinia virus as a cloning vector. Based on cloned coronaviral cDNA, we describe the generation of recombinant coronaviruses and stable cell lines containing coronaviral replicon RNAs. Initially, the vaccinia virus-based reverse genetic system was established for the generation of recombinant human coronavirus 229E. However, it is also applicable to the generation of other coronaviruses, such as the avian infectious bronchitis virus, mouse hepatitis virus, and SARS coronavirus.