Demonstration of bacterium‐free very rapid reverse genetics system using mumps virus. (2nd November 2022)

• Record Type: Journal Article
• Authors: Bae, Chaewon; Katoh, Hiroshi; Wakata, Aika; Sakata, Masafumi; Kato, Fumihiro; Takeda, Makoto
• Is Part Of: Microbiology and immunology. Volume 67:Number 1(2023)
• Page Start: 44
• Page End: 47
• Publication Date: 2022-11-02
• Subjects: Escherichia coli‐free -- mumps virus -- OriCiro cell‐free cloning system -- PCR -- reverse genetics; Microbiology -- Periodicals; Immunology -- Periodicals; Allergy and Immunology -- Periodicals; Microbiology -- Periodicals; Microbiologie -- Périodiques; Immunologie -- Périodiques; 579
• DOI: 10.1111/1348-0421.13032
• Languages: English
• ISSNs: 0385-5600

Abstract

Abstract: The reverse genetics system is a very powerful tool for analyzing the molecular mechanisms of viral propagation and pathogenesis. However, full‐length genome plasmid construction is highly time‐consuming and laborious, and undesired mutations may be introduced by Escherichia coli . This study shows a very rapid E. coli ‐free method of full‐genome construction using the mumps virus as an example. This method was able to reduce dramatically the time for full‐genome construction, which was used very efficiently for virus rescue, from several days or more to ~2 days, with a similar accuracy and yield to the conventional method using E. coli /plasmid.