Generation of Full-Length Infectious cDNA Clones of Middle East Respiratory Syndrome Coronavirus
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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 virusspecific 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 patientisolated 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.
Abstract
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 virusspecific 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 patientisolated 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.
참고문헌 (54)
- Alexandersen S, "Middle East respiratory syndrome coronavirus antibody reactors among camels in Dubai, United Arab Emirates, in 2005", Transbound. Emerg. Dis, 61, pp.105~108 (2014)
- Haagmans BL, "Middle East respiratory syndrome coronavirus in dromedary camels : an outbreak investigation", Lancet Infect. Dis, 14, pp.140~145 (2014)
- Zaki AM, "Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia", N. Engl. J. Med, 367, pp.1814~1820 (2012)
- Azhar EI, "Evidence for camel-to-human transmission of MERS coronavirus", N. Engl. J. Med, 370, pp.2499~2505 (2014)
- Ferguson NM, "Identification of MERS-CoV in dromedary camels", Lancet Infect. Dis, 14, pp.93~94 (2014)
- Kim KH, "Middle East respiratory syndrome coronavirus(MERS-CoV)outbreak in South Korea, 2015 : epidemiology, characteristics and public health implications", J. Hosp. Infect, 95, pp.207~213 (2017)
- Ki M, "2015 MERS outbreak in Korea: hospital-tohospital transmission", Epidemiol. Health, 37, pp.e2015033~ (2015)
- Kim SW, "Risk factors for transmission of Middle East respiratory syndrome coronavirus infection during the 2015 outbreak in South Korea", Clin. Infect. Dis, 64, pp.551~557 (2017)
- Lim PL, "Middle East respiratory syndrome (MERS) in Asia: lessons gleaned from the South Korean outbreak", Trans. R. Soc. Trop. Med. Hyg, 109, pp.541~542 (2015)
- Chowell G, "Transmission characteristics of MERS and SARS in the healthcare setting : a comparative study", BMC Med, 13, pp.210~ (2015)
- Fung IC, "Middle East respiratory syndrome in the Republic of Korea: transparency and communication are key", Western Pac. Surveill. Response J, 6, pp.1~2 (2015)
- Sherertz RJ, "A cloud adult: the Staphylococcus aureus-virus interaction revisited", Ann. Intern. Med, 124, pp.539~547 (1996)
- Almazan F, "Engineering a replicationcompetent, propagation-defective Middle East respiratory syndrome coronavirus as a vaccine candidate", MBio, 4, pp.e00650~e00613 (2013)
- Scobey T, "Reverse genetics with a fulllength infectious cDNA of the Middle East respiratory syndrome coronavirus", Proc. Natl. Acad. Sci. USA, 110, pp.16157~16162 (2013)
- 전형태, "Kaposi’s Sarcoma-Associated Herpesvirus Infection Modulates the Proliferation of Glioma Stem-Like Cells", Journal of Microbiology and Biotechnology, 28(1), pp.165~174 (2018), 한국미생물·생명공학회
- 박병주, "Pathogenesis of Human Norovirus Genogroup II Genotype 4 in Post- Weaning Gnotobiotic Pigs", Journal of Microbiology and Biotechnology, 28(12), pp.2133~2140 (2018), 한국미생물·생명공학회
- 윤성진, "Optimized Expression, Purification, and Rapid Detection of Recombinant Influenza Nucleoproteins Expressed in Sf9 Insect Cells", Journal of Microbiology and Biotechnology, 28(10), pp.1683~1690 (2018), 한국미생물·생명공학회
- Kang HS, "Transgenic expression of non-structural genes of Theiler’s virus suppresses initial viral replication and pathogenesis of demyelination", J. Neuroinflammation, 13, pp.133~ (2016)
- 강상민, "Hepatitis E Virus Methyltransferase Inhibits Type I Interferon Induction by Targeting RIG-I", Journal of Microbiology and Biotechnology, 28(9), pp.1554~1562 (2018), 한국미생물·생명공학회
- 강상민, "Host Innate Immunity against Hepatitis E Virus and Viral Evasion Mechanisms", Journal of Microbiology and Biotechnology, 27(10), pp.1727~1735 (2017), 한국미생물·생명공학회
- 강상민, "Primary lymphocyte infection models for KSHV and its putative tumorigenesis mechanisms in B cell lymphomas", The Journal of Microbiology, 55(5), pp.319~329 (2017), 한국미생물학회
- 김은하, "Hepatitis E Virus Papain-Like Cysteine Protease Inhibits Type I Interferon Induction by Down-Regulating Melanoma Differentiation-Associated Gene 5", Journal of Microbiology and Biotechnology, 28(11), pp.1908~1915 (2018), 한국미생물·생명공학회
- Gholamalipour Y, "3’ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character-RNASeq analyses", Nucleic Acids Res, 46, pp.9253~9263 (2018)
- Racaniello VR, "Studying poliovirus with infectious cloned cDNA", Rev. Infect. Dis, 6(Suppl 2), pp.S514~e515 (1984)
- Racaniello VR, "Cloned poliovirus complementary DNA is infectious in mammalian cells", Science, 214, pp.916~919 (1981)
- Stobart CC, "RNA virus reverse genetics and vaccine design", Viruses, 6, pp.2531~2550 (2014)
- Matsui SM, "The isolation and characterization of a Norwalk virus-specific cDNA", J. Clin. Invest, 87, pp.1456~1461 (1991)
- Katayama K, "Plasmid-based human norovirus reverse genetics system produces reporter-tagged progeny virus containing infectious genomic RNA", Proc. Natl. Acad. Sci. USA, 111, pp.E4043~E4052 (2014)
- Panda SK, "The in vitro-synthesized RNA from a cDNA clone of hepatitis E virus is infectious", J. Virol, 74, pp.2430~2437 (2000)
- Shan C, "An infectious cDNA clone of Zika virus to study viral virulence, mosquito transmission, and antiviral inhibitors", Cell Host Microbe, 19, pp.891~900 (2016)
- Tsetsarkin KA, "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 (2016)
- Yang Y, "A cDNA clonelaunched platform for high-yield production of inactivated Zika vaccine", EBioMedicine, 17, pp.145~156 (2017)
- Zhang L, "Tet-inducible production of infectious Zika virus from the full-length cDNA clones of african- and asian-lineage strains", Viruses, 10 (2018)
- Yount B, "Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus", Proc. Natl. Acad. Sci. USA, 100, pp.12995~13000 (2003)
- Almazan F, "Engineering infectious cDNAs of coronavirus as bacterial artificial chromosomes", Methods Mol. Biol, 1282, pp.135~152 (2015)
- Cockrell AS, "Efficient reverse genetic systems for rapid genetic manipulation of emergent and preemergent infectious coronaviruses", Methods Mol. Biol, 1602, pp.59~81 (2017)
- Boehme KW, "Reverse genetics for mammalian reovirus", Methods, 55, pp.109~113 (2011)
- Roner MR, "Reovirus reverse genetics : Incorporation of the CAT gene into the reovirus genome", Proc. Natl. Acad. Sci. USA, 98, pp.8036~8041 (2001)
- de Wit E, "Efficient generation and growth of influenza virus A", PR, 34 from eight cDNA fragments, pp.Virus Res, 2004[DOI]
- Jackson D, "A reverse genetics approach for recovery of recombinant influenza B viruses entirely from cDNA", J. Virol, 76, pp.11744~11747 (2002)
- Flatz L, "Recovery of an arenavirus entirely from RNA polymerase I", II-driven cDNA, 2006, pp.103, 4663~4668
- Ndung'u T, "Construction and analysis of an infectious human Immunodeficiency virus type 1 subtype C molecular clone", J. Virol, 75, pp.4964~4972 (2001)
- Prakash K, "Isolation and characterization of an infectious molecular clone of the MN strain of HIV-1", Biochem. Biophys. Res. Commun, 179, pp.1377~1383 (1991)
- Martinez-Sobrido L, "Reporterexpressing, replicating-competent recombinant arenaviruses", Viruse, 8 (2016)
- Castrucci MR, "Attenuation of influenza A virus by insertion of a foreign epitope into the neuraminidase", J. Virol, 66, pp.4647~4653 (1992)
- Halfmann P, "Replication-deficient ebolavirus as a vaccine candidate", J. Virol, 83, pp.3810~3815 (2009)
- Gibson DG, "Enzymatic assembly of DNA molecules up to several hundred kilobases", Nat. Methods, 6, pp.343~345 (2009)
- Gibson DG, "Enzymatic assembly of overlapping DNA fragments", Methods Enzymol, 498, pp.349~361 (2011)
- Fernandez-Rodriguez J, "Engineering RGB color vision into Escherichia coli", Nat. Chem. Biol, 13, pp.706~708 (2017)
- Song M, "Antioxidant defense by thioredoxin can occur independently of canonical thiol-disulfide oxidoreductase enzymatic activity", Cell Rep, 14, pp.2901~2911 (2016)
- Song M, "Control of type III protein secretion using a minimal genetic system", Nat. Commun, 8, pp.14737~ (2017)
- Dudek T, "Replication-defective viruses as vaccines and vaccine vectors", Virology, 344, pp.230~239 (2006)
- Giel-Moloney M, "A novel approach to a rabies vaccine based on a recombinant single-cycle flavivirus vector", Vaccine, 35, pp.6898~6904 (2017)
- Welch SK, "Construction and evaluation of genetically engineered replication-defective porcine reproductive and respiratory syndrome virus vaccine candidates", Vet. Immunol. Immunopathol, 102, pp.277~290 (2004)