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Demonstration of bacterium‐free very rapid reverse genetics system using mumps virus
Author(s) -
Bae Chaewon,
Katoh Hiroshi,
Wakata Aika,
Sakata Masafumi,
Kato Fumihiro,
Takeda Makoto
Publication year - 2023
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/1348-0421.13032
Subject(s) - reverse genetics , biology , plasmid , mumps virus , genome , virus , escherichia coli , genetics , virology , gene , computational biology
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.