生物
翻译(生物学)
内部核糖体进入位点
重组DNA
核糖核酸
遗传学
病毒学
计算生物学
反向遗传学
基因组
基因
信使核糖核酸
作者
Tomokazu Tamura,Hirotaka Yamamoto,Saho Ogino,Yuhei Morioka,Shuhei Tsujino,Rigel Suzuki,Takahiro Hiono,Saori Suzuki,Norikazu Isoda,Yoshihiro Sakoda,Takasuke Fukuhara
摘要
The lack of versatility of reverse genetics systems remains a bottleneck in viral research. Especially when (re-)emerging viruses reach pandemic levels, rapid characterization and establishment of effective countermeasures using recombinant viruses are beneficial in disease control. Indeed, numerous studies have attempted to establish and improve the methods. The circular polymerase extension reaction (CPER) method has overcome major obstacles in generating recombinant viruses. However, this method has not yet been examined for positive-strand RNA viruses that use cap-independent, internal ribosome entry site-mediated translation. Here, we engineered a suitable gene cassette to expand the CPER method for all positive-strand RNA viruses. Furthermore, we overcame the difficulty of generating recombinant viruses because of low transfection efficiency. Using this modified method, we also successfully generated reporter viruses and recombinant viruses from a field sample without virus isolation. Taking these findings together, our adapted methodology is an innovative technology that could help advance virologic research.
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