Author(s)

Jiapeng Luo ¹, Ping Zhao ²

Affiliation(s)

¹ School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai, 200433, China
² Department of Biomedical Defense, Faculty of Naval Medicine, Naval Medical University, Shanghai, 200433, China

Corresponding Author

Ping Zhao

ABSTRACT

Chikungunya virus (CHIKV) is the pathogen that causes Chikungunya fever, and there is no specific antiviral drug treatment for CHIKV infection. A simple and efficient CHIKV infectious clone is an important tool for studying the mechanism of CHIKV infection and development of vaccine and drug. Based on the cDNA sequence of the virus published in GenBank, we obtained the whole sequence of the viral genome by cleavage and ligation of adjacent fragments, and inserted a mammalian intron sequence which does not affect the production of the virus to ensure the stable inheritance of the virus. We introduced an EcoR Ⅰ restriction endonuclease cleavage site and a T7 promoter sequence at the 5' end, and introduced a Not I restriction endonuclease cleavage site at the 3' end. Then the entire sequence was inserted into pVRC-L vector to obtain the infectious clone pCHIKV. The transfected supernatant was obtained by transcribing and transfecting into BHK-21 cells. The virus was identified by sequencing of the transfected supernatant, empty plaque, indirect immunofluorescence and mice infection test. The results showed that pCHIKV and the transfected supernatant were sequenced as expected, the viral titer was calculated as 5*10⁷ FFU/ml, the virus could infect Vero E6 cells, the virus-infected mice became ill on the 3rd day, and all of them died on the 4th day, and the virus were present in the liver tissues of the dead mice. These results indicated the successful construction of the CHIKV infected clone.

KEYWORDS

Chikungunya virus, infectious clones, reverse genetics

CITE THIS PAPER

Jiapeng Luo, Ping Zhao, Construction and Characterization of an Infectious Clone of Chikungunya Virus Strain 37997. MEDS Basic Medicine (2024) Vol. 2: 39-47. DOI: http://dx.doi.org/10.23977/medbm.2024.020106.

REFERENCES

[1] Ross, R. W. (1956). The Newala epidemic. III. The virus: isolation, pathogenic properties and relationship to the epidemic. J Hyg (Lond) 54, 177-191. 
[2] Robinson, M. C. (1955). An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952-53. I. Clinical features. Trans R Soc Trop Med Hyg 49, 28-32.
[3] Lumsden, W. H. (1955). An epidemic of virus disease in Southern Province, Tanganyika Territory, in 1952-53. II. General description and epidemiology. Trans R Soc Trop Med Hyg 49, 33-57.
[4] de Lima Cavalcanti, T. Y. V., Pereira, M. R., de Paula, S. A.-O. & Franca, R. A.-O. (2022). A Review on Chikungunya Virus Epidemiology, Pathogenesis and Current Vaccine Development. Virus 14(5):969.
[5] Suhrbier, A. A.-O. (2019). Rheumatic manifestations of chikungunya: emerging concepts and interventions. Nat Rev Rheumatol 15(10):597-611.
[6] Liu, L. B.Li, M.Gao N.Shen, J. Y.Shen, Z. Y.Fan, D. Y. (2022). Epidemiological and clinical characteristics of the chikungunya outbreak in Ruili City, Yunnan Province, China. J Med Virol 94(2):499-506.
[7] Pan J Fau. Fang, C. Yan, H. Zhan, B, d. Sun, Y. Liu, Y. (2019). Chikungunya Fever Outbreak, Zhejiang Province, China, 2017. Emerg Infect Dis 25(8):1589-1591.
[8] Khan, A. H. Morita, K. Parquet, M.D.C. Hasebe, F. Mathenge, E.G.M. Igarashi, A. (2022). Complete nucleotide sequence of chikungunya virus and evidence for an internal polyadenylation site. J Gen Virol 83(Pt 12):3075-3084.
[9] Lescar, J.Roussel, A. Wien, M,W. Navaza, J. Fuller, S, D. Wengler, G. Rey, F.A. (2001). The Fusion glycoprotein shell of Semliki Forest virus: an icosahedral assembly primed for fusogenic activation at endosomal pH. Cell 105(1):137-148.
[10] Couderc, T. Chretien, F. Schilte, C. Disson, O. Brigitte, M. Guivel-Benhassine, F. (2022). A mouse model for Chikungunya: young age and inefficient type-I interferon signaling are risk factors for severe disease. PLoS Pathog 4(2):e29.
[11] Volk, S. M. Chen, R. Tsetsarkin, K.A. Adams, A, P. Garcia, T, I. Sall, A, A. (2022). Genome-scale phylogenetic analyses of chikungunya virus reveal independent emergences of recent epidemics and various evolutionary rates. J Virol 4(13):6497-6504.
[12] Tsetsarkin, K. A., Vanlandingham Dl Fau - McGee, C. E., McGee Ce Fau - Higgs, S. & Higgs, S. (2007). A single mutation in chikungunya virus affects vector specificity and epidemic potential. PLoS Pathog 3(12):e201.

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