Reverse Genetics of Zika Virus
We have established a reverse genetic system for Zika virus (ZIKV). Five shuttle plasmids were constructed and assembled into the full-length cDNA clone of ZIKV genome. To ensure the stability of the cDNA clone, we used a low copy vector (pACYC177) and a set of unique restriction enzyme sites on the ZIKV genome to assemble the full-length cDNA clone. A T7 promoter was engineered in front of the viral 5′ UTR for in vitro transcription. A hepatitis delta virus ribozyme (HDVr) sequence was engineered following the viral 3′ UTR for generation of the authentic 3′ end of the RNA transcript.
Key wordsFlavivirus Zika Mosquito Dengue West Nile Guillain-Barré syndrome Microcephaly
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