Advertisement

Reverse Genetics of Zika Virus

  • Chao Shan
  • Xuping Xie
  • Pei-Yong ShiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1602)

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.

Key words

Flavivirus Zika Mosquito Dengue West Nile Guillain-Barré syndrome Microcephaly 

References

  1. 1.
    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 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    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. 712–746Google Scholar
  3. 3.
    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 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    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)Google Scholar
  5. 5.
    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 2:170–172CrossRefPubMedGoogle Scholar
  6. 6.
    Dick GW, Kitchen SF, Haddow AJ (1952) Zika virus. I. Isolations and serological specificity. Trans R Soc Trop Med Hyg 46(5):509–520CrossRefPubMedGoogle Scholar
  7. 7.
    Petersen LR, Jamieson DJ, Powers AM, Honein MA (2016) Zika Virus. N Engl J Med. doi: 10.1056/NEJMra1602113 Google Scholar
  8. 8.
    Fauci AS, Morens DM (2016) Zika virus in the Americas—yet another arbovirus threat. N Engl J Med. doi: 10.1056/NEJMp1600297 Google Scholar
  9. 9.
    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):5847–5856CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    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 CrossRefPubMedGoogle Scholar
  11. 11.
    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 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Departments of Biochemistry & Molecular Biology and Pharmacology & Toxicology, and Sealy Center for Structural Biology & Molecular BiophysicsUniversity of Texas Medical BranchGalvestonUSA

Personalised recommendations