Abstract
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.
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References
Lanciotti RS, Kosoy OL, Laven JJ, Velez JO, Lambert AJ, Johnson AJ, Stanfield SM, Duffy MR (2008) Genetic and serologic properties of Zika virus associated with an epidemic, Yap State, Micronesia, 2007. Emerg Infect Dis 14(8):1232–1239. doi:10.3201/eid1408.080287
Lindenbach BD, Murray CJ, Thiel H-J, Rice CM (2013) Flaviviridae In D.M. Knipe and P.M. Howley (ed), Fields virology, 6th edn, vol 1. Lippincott William & Wilkins, Philadelphia, Pap. 712–746
Musso D, Roche C, Robin E, Nhan T, Teissier A, Cao-Lormeau VM (2015) Potential sexual transmission of Zika virus. Emerg Infect Dis 21(2):359–361. doi:10.3201/eid2102.141363
Musso D, Nhan T, Robin E, Roche C, Bierlaire D, Zisou K, Shan Yan A, Cao-Lormeau VM, Broult J (2014) Potential for Zika virus transmission through blood transfusion demonstrated during an outbreak in French Polynesia, November 2013 to February 2014. Euro Surveill 19(14)
Shan C, Xie X, Barrett ADT, Garcia-Blanco MA, Tesh RB, Vasconcelos PFDC, Vasilakis N, Weaver SC, Shi PY (2016) Zika virus: diagnosis, therapeutics, and vaccine. ACS Infect Dis 2:170–172
Dick GW, Kitchen SF, Haddow AJ (1952) Zika virus. I. Isolations and serological specificity. Trans R Soc Trop Med Hyg 46(5):509–520
Petersen LR, Jamieson DJ, Powers AM, Honein MA (2016) Zika Virus. N Engl J Med. doi:10.1056/NEJMra1602113
Fauci AS, Morens DM (2016) Zika virus in the Americas—yet another arbovirus threat. N Engl J Med. doi:10.1056/NEJMp1600297
Shi PY, Tilgner M, Lo MK, Kent KA, Bernard KA (2002) Infectious cDNA clone of the epidemic West Nile virus from New York City. J Virol 76(12):5847–5856
Zou G, Xu HY, Qing M, Wang QY, Shi PY (2011) Development and characterization of a stable luciferase dengue virus for high-throughput screening. Antivir Res 91(1):11–19. doi:10.1016/j.antiviral.2011.05.001
Li XD, Li XF, Ye HQ, Deng CL, Ye Q, Shan C, Shang BD, Xu LL, Li SH, Cao SB, Yuan ZM, Shi PY, Qin CF, Zhang B (2014) Recovery of a chemically synthesized Japanese encephalitis virus reveals two critical adaptive mutations in NS2B and NS4A. J Gen Virol 95(Pt 4):806–815. doi:10.1099/vir.0.061838-0
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Shan, C., Xie, X., Shi, PY. (2017). Reverse Genetics of Zika Virus. In: Perez, D. (eds) Reverse Genetics of RNA Viruses. Methods in Molecular Biology, vol 1602. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6964-7_4
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DOI: https://doi.org/10.1007/978-1-4939-6964-7_4
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