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Generation of Full-Length Infectious cDNA Clones of Middle East Respiratory Syndrome Coronavirus

  • Lee, Jeong Yoon (Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University) ;
  • Bae, Sojung (Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University) ;
  • Myoung, Jinjong (Korea Zoonosis Research Institute and Genetic Engineering Research Institute, Chonbuk National University)
  • 투고 : 2019.05.07
  • 심사 : 2019.06.01
  • 발행 : 2019.06.28

초록

Middle East respiratory syndrome coronavirus (MERS-CoV) was first identified in Saudi Arabia in 2012 and related infection cases have been reported in over 20 countries. Roughly 10,000 human cases have so far been reported in total with fatality rates at up to 40%. The majority of cases have occurred in Saudi Arabia with mostly sporadic outbreaks outside the country except for the one in South Korea in 2015. The Korean MERS-CoV strain was isolated from the second Korean patient and its genome was fully sequenced and deposited. To develop virus-specific protective and therapeutic agents against the Korean isolate and to investigate molecular determinants of virus-host interactions, it is of paramount importance to generate its full-length cDNA. Here we report that two full-length cDNAs from a Korean patient-isolated MERS-CoV strain were generated by a combination of conventional cloning techniques and efficient Gibson assembly reactions. The full-length cDNAs were validated by restriction analysis and their sequence was verified by Sanger method. The resulting cDNA was efficiently transcribed in vitro and the T7 promoter-driven expression was robust. The resulting reverse genetic system will add to the published list of MERS-CoV cDNAs and facilitate the development of Korean isolate-specific antiviral measures.

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참고문헌

  1. Alexandersen S, Kobinger GP, Soule G, Wernery U. 2014. Middle East respiratory syndrome coronavirus antibody reactors among camels in Dubai, United Arab Emirates, in 2005. Transbound. Emerg. Dis. 61: 105-108. https://doi.org/10.1111/tbed.12212
  2. Haagmans BL, Al Dhahiry SH, Reusken CB, Raj VS, Galiano M, Myers R, et al. 2014. Middle East respiratory syndrome coronavirus in dromedary camels: an outbreak investigation. Lancet Infect. Dis. 14: 140-145. https://doi.org/10.1016/S1473-3099(13)70690-X
  3. Zaki AM, van Boheemen S, Bestebroer TM, Osterhaus AD, Fouchier RA. 2012. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N. Engl. J. Med. 367: 1814-1820. https://doi.org/10.1056/NEJMoa1211721
  4. Azhar EI, El-Kafrawy SA, Farraj SA, Hassan AM, Al-Saeed MS, Hashem AM, et al. 2014. Evidence for camel-to-human transmission of MERS coronavirus. N. Engl. J. Med. 370: 2499-2505. https://doi.org/10.1056/NEJMoa1401505
  5. Ferguson NM, Van Kerkhove MD. 2014. Identification of MERS-CoV in dromedary camels. Lancet Infect. Dis. 14: 93-94. https://doi.org/10.1016/S1473-3099(13)70691-1
  6. Kim KH, Tandi TE, Choi JW, Moon JM, Kim MS. 2017. Middle East respiratory syndrome coronavirus (MERS-CoV) outbreak in South Korea, 2015: epidemiology, characteristics and public health implications. J. Hosp. Infect. 95: 207-213. https://doi.org/10.1016/j.jhin.2016.10.008
  7. Ki M. 2015. 2015 MERS outbreak in Korea: hospital-tohospital transmission. Epidemiol. Health. 37: e2015033. https://doi.org/10.4178/epih/e2015033
  8. Kim SW, Park JW, Jung HD, Yang JS, Park YS, Lee C, et al. 2017. Risk factors for transmission of Middle East respiratory syndrome coronavirus infection during the 2015 outbreak in South Korea. Clin. Infect. Dis. 64: 551-557. https://doi.org/10.1093/cid/ciw768
  9. Lim PL. 2015. Middle East respiratory syndrome (MERS) in Asia: lessons gleaned from the South Korean outbreak. Trans. R. Soc. Trop. Med. Hyg. 109: 541-542. https://doi.org/10.1093/trstmh/trv064
  10. Chowell G, Abdirizak F, Lee S, Lee J, Jung E, Nishiura H, et al. 2015. Transmission characteristics of MERS and SARS in the healthcare setting: a comparative study. BMC Med. 13: 210. https://doi.org/10.1186/s12916-015-0450-0
  11. Fung IC, Tse ZT, Chan BS, Fu KW. 2015. Middle East respiratory syndrome in the Republic of Korea: transparency and communication are key. Western Pac. Surveill. Response J. 6: 1-2. https://doi.org/10.5365/wpsar.2015.6.2.011
  12. Sherertz RJ, Reagan DR, Hampton KD, Robertson KL, Streed SA, Hoen HM, et al. 1996. A cloud adult: the Staphylococcus aureus-virus interaction revisited. Ann. Intern. Med. 124: 539-547. https://doi.org/10.7326/0003-4819-124-6-199603150-00001
  13. Almazan F, DeDiego ML, Sola I, Zuniga S, Nieto-Torres JL, Marquez-Jurado S, et al. 2013. Engineering a replicationcompetent, propagation-defective Middle East respiratory syndrome coronavirus as a vaccine candidate. MBio. 4: e00650-00613.
  14. Scobey T, Yount BL, Sims AC, Donaldson EF, Agnihothram SS, Menachery VD, et al. 2013. Reverse genetics with a fulllength infectious cDNA of the Middle East respiratory syndrome coronavirus. Proc. Natl. Acad. Sci. USA 110: 16157-16162. https://doi.org/10.1073/pnas.1311542110
  15. Jeon H, Kang YH, Yoo SM, Park MJ, Park JB, Lee SH, et al. 2018. Kaposi's sarcoma-associated herpesvirus infection modulates the proliferation of glioma stem-like cells. J. Microbiol. Biotechnol. 28: 165-174. https://doi.org/10.4014/jmb.1709.09001
  16. Park BJ, Jung ST, Choi CS, Myoung J, Ahn HS, Han SH, et al. 2018. Pathogenesis of human norovirus genogroup II genotype 4 in post-weaning gnotobiotic pigs. J. Microbiol. Biotechnol. 28: 2133-2140. https://doi.org/10.4014/jmb.1810.09061
  17. Yoon SJ, Park YJ, Kim HJ, Jang J, Lee SJ, Koo S, et al. 2018. Optimized expression, purification, and rapid detection of recombinant influenza nucleoproteins expressed in Sf9 insect cells. J. Microbiol. Biotechnol. 28: 1683-1690. https://doi.org/10.4014/jmb.1805.05053
  18. Kang HS, Myoung J, So EY, Bahk YY, Kim BS. 2016. Transgenic expression of non-structural genes of Theiler's virus suppresses initial viral replication and pathogenesis of demyelination. J. Neuroinflammation 13: 133. https://doi.org/10.1186/s12974-016-0597-4
  19. Kang S, Choi C, Choi I, Han KN, Rho SW, Choi J, et al. 2018. Hepatitis E virus methyltransferase inhibits type i interferon induction by targeting RIG-I. J. Microbiol. Biotechnol. 28: 1554-1562. https://doi.org/10.4014/jmb.1808.08058
  20. Kang S, Myoung J. 2017. Host innate immunity against hepatitis E virus and viral evasion mechanisms. J. Microbiol. Biotechnol. 27: 1727-1735. https://doi.org/10.4014/jmb.1708.08045
  21. Kang S, Myoung J. 2017. Primary lymphocyte infection models for KSHV and its putative tumorigenesis mechanisms in B cell lymphomas. J. Microbiol. 55: 319-329. https://doi.org/10.1007/s12275-017-7075-2
  22. Kim E, Myoung J. 2018. Hepatitis E virus papain-like cysteine protease inhibits type i interferon induction by down-regulating melanoma differentiation-associated gene 5. J. Microbiol. Biotechnol. 28: 1908-1915. https://doi.org/10.4014/jmb.1809.09028
  23. Gholamalipour Y, Karunanayake Mudiyanselage A, Martin CT. 2018. 3' end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character-RNASeq analyses. Nucleic Acids Res. 46: 9253-9263. https://doi.org/10.1093/nar/gky796
  24. Racaniello VR. 1984. Studying poliovirus with infectious cloned cDNA. Rev. Infect. Dis. 6 Suppl 2: S514-515. https://doi.org/10.1093/clinids/6.Supplement_2.S514
  25. Racaniello VR, Baltimore D. 1981. Cloned poliovirus complementary DNA is infectious in mammalian cells. Science 214: 916-919. https://doi.org/10.1126/science.6272391
  26. Stobart CC, Moore ML. 2014. RNA virus reverse genetics and vaccine design. Viruses 6: 2531-2550. https://doi.org/10.3390/v6072531
  27. Matsui SM, Kim JP, Greenberg HB, Su W, Sun Q, Johnson PC, et al. 1991. The isolation and characterization of a Norwalk virus-specific cDNA. J. Clin. Invest. 87: 1456-1461. https://doi.org/10.1172/JCI115152
  28. Katayama K, Murakami K, Sharp TM, Guix S, Oka T, Takai-Todaka R, et al. 2014. Plasmid-based human norovirus reverse genetics system produces reporter-tagged progeny virus containing infectious genomic RNA. Proc. Natl. Acad. Sci. USA 111: E4043-4052. https://doi.org/10.1073/pnas.1415096111
  29. Panda SK, Ansari IH, Durgapal H, Agrawal S, Jameel S. 2000. The in vitro-synthesized RNA from a cDNA clone of hepatitis E virus is infectious. J. Virol. 74: 2430-2437. https://doi.org/10.1128/JVI.74.5.2430-2437.2000
  30. Shan C, Xie X, Muruato AE, Rossi SL, Roundy CM, Azar SR, et al. 2016. An infectious cDNA clone of Zika virus to study viral virulence, mosquito transmission, and antiviral inhibitors. Cell Host Microbe 19: 891-900. https://doi.org/10.1016/j.chom.2016.05.004
  31. Tsetsarkin KA, Kenney H, Chen R, Liu G, Manukyan H, Whitehead SS, et al. 2016. A full-length infectious cDNA clone of Zika virus from the 2015 epidemic in Brazil as a genetic platform for studies of virus-host interactions and vaccine development. MBio 7.
  32. Yang Y, Shan C, Zou J, Muruato AE, Bruno DN, de Almeida Medeiros Daniele B, et al. 2017. A cDNA clonelaunched platform for high-yield production of inactivated Zika vaccine. EBioMedicine 17: 145-156. https://doi.org/10.1016/j.ebiom.2017.02.003
  33. Zhang L, Ji W, Lyu S, Qiao L, Luo G. 2018. Tet-inducible production of infectious Zika virus from the full-length cDNA clones of african- and asian-lineage strains. Viruses 10.
  34. Yount B, Curtis KM, Fritz EA, Hensley LE, Jahrling PB, Prentice E, et al. 2003. Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. Proc. Natl. Acad. Sci. USA 100: 12995-13000. https://doi.org/10.1073/pnas.1735582100
  35. Almazan F, Marquez-Jurado S, Nogales A, Enjuanes L. 2015. Engineering infectious cDNAs of coronavirus as bacterial artificial chromosomes. Methods Mol. Biol. 1282: 135-152. https://doi.org/10.1007/978-1-4939-2438-7_13
  36. Cockrell AS, Beall A, Yount B, Baric R. 2017. Efficient reverse genetic systems for rapid genetic manipulation of emergent and preemergent infectious coronaviruses. Methods Mol. Biol. 1602: 59-81. https://doi.org/10.1007/978-1-4939-6964-7_5
  37. Boehme KW, Ikizler M, Kobayashi T, Dermody TS. 2011. Reverse genetics for mammalian reovirus. Methods 55: 109-113. https://doi.org/10.1016/j.ymeth.2011.07.002
  38. Roner MR, Joklik WK. 2001. Reovirus reverse genetics: Incorporation of the CAT gene into the reovirus genome. Proc. Natl. Acad. Sci. USA 98: 8036-8041. https://doi.org/10.1073/pnas.131203198
  39. de Wit E, Spronken MI, Bestebroer TM, Rimmelzwaan GF, Osterhaus AD, Fouchier RA. 2004. Efficient generation and growth of influenza virus A/PR/8/34 from eight cDNA fragments. Virus Res. 103: 155-161. https://doi.org/10.1016/j.virusres.2004.02.028
  40. Jackson D, Cadman A, Zurcher T, Barclay WS. 2002. A reverse genetics approach for recovery of recombinant influenza B viruses entirely from cDNA. J. Virol. 76: 11744-11747. https://doi.org/10.1128/JVI.76.22.11744-11747.2002
  41. Flatz L, Bergthaler A, de la Torre JC, Pinschewer DD. 2006. Recovery of an arenavirus entirely from RNA polymerase I/II-driven cDNA. Proc. Natl. Acad. Sci. USA 103: 4663-4668. https://doi.org/10.1073/pnas.0600652103
  42. Ndung'u T, Renjifo B, Essex M. 2001. Construction and analysis of an infectious human Immunodeficiency virus type 1 subtype C molecular clone. J. Virol. 75: 4964-4972. https://doi.org/10.1128/JVI.75.11.4964-4972.2001
  43. Prakash K, Hodinka RL, Hullihen DM, Plotkin SA. 1991. Isolation and characterization of an infectious molecular clone of the MN strain of HIV-1. Biochem. Biophys. Res. Commun. 179: 1377-1383. https://doi.org/10.1016/0006-291X(91)91725-R
  44. Martinez-Sobrido L, de la Torre JC. 2016. Reporterexpressing, replicating-competent recombinant arenaviruses. Viruse 8.
  45. Castrucci MR, Bilsel P, Kawaoka Y. 1992. Attenuation of influenza A virus by insertion of a foreign epitope into the neuraminidase. J. Virol. 66: 4647-4653. https://doi.org/10.1128/JVI.66.8.4647-4653.1992
  46. Halfmann P, Ebihara H, Marzi A, Hatta Y, Watanabe S, Suresh M, et al. 2009. Replication-deficient ebolavirus as a vaccine candidate. J. Virol. 83: 3810-3815. https://doi.org/10.1128/JVI.00074-09
  47. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA, 3rd, Smith HO. 2009. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6: 343-345. https://doi.org/10.1038/nmeth.1318
  48. Gibson DG. 2011. Enzymatic assembly of overlapping DNA fragments. Methods Enzymol. 498: 349-361. https://doi.org/10.1016/B978-0-12-385120-8.00015-2
  49. Fernandez-Rodriguez J, Moser F, Song M, Voigt CA. 2017. Engineering RGB color vision into Escherichia coli. Nat. Chem. Biol. 13: 706-708. https://doi.org/10.1038/nchembio.2390
  50. Song M, Kim JS, Liu L, Husain M, Vazquez-Torres A. 2016. Antioxidant defense by thioredoxin can occur independently of canonical thiol-disulfide oxidoreductase enzymatic activity. Cell Rep. 14: 2901-2911. https://doi.org/10.1016/j.celrep.2016.02.066
  51. Song M, Sukovich DJ, Ciccarelli L, Mayr J, Fernandez-Rodriguez J, Mirsky EA, et al. 2017. Control of type III protein secretion using a minimal genetic system. Nat. Commun. 8: 14737. https://doi.org/10.1038/ncomms14737
  52. Dudek T, Knipe DM. 2006. Replication-defective viruses as vaccines and vaccine vectors. Virology 344: 230-239. https://doi.org/10.1016/j.virol.2005.09.020
  53. Giel-Moloney M, Rumyantsev AA, David F, Figueiredo M, Feilmeier B, Mebatsion T, et al. 2017. A novel approach to a rabies vaccine based on a recombinant single-cycle flavivirus vector. Vaccine 35: 6898-6904. https://doi.org/10.1016/j.vaccine.2017.08.055
  54. Welch SK, Jolie R, Pearce DS, Koertje WD, Fuog E, Shields SL, et al. 2004. Construction and evaluation of genetically engineered replication-defective porcine reproductive and respiratory syndrome virus vaccine candidates. Vet. Immunol. Immunopathol. 102: 277-290. https://doi.org/10.1016/j.vetimm.2004.09.022

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