Objective To express severe fever with thrombocytopenia syndrome virus (SFTSV) glycoprotein Gc in prokaryotic cells, and to preliminarily study the immunogenicity of glycoprotein Gc. Methods The DNA sequence encoding envelope glycoprotein Gc in SFTSV HB29 strain was chemically synthesized after codon optimization based on Escherichia coli codon preference. The expression vector pET-His-Gc was constructed by cloning the target gene into the vector pET-His. The sequence of the vector was confirmed by sequencing. Protein expression was carried out by transforming the vector into BL21 (DE3) competent cells. Target protein was purified and verified by SDS-PAGE and Western blot. Purified protein was used to immunize New Zealand white rabbits. The rabbit sera were collected for determination of antibody titer. Results The expression vector pET-His-Gc was successful constructed. Glycoprotein Gc was expressed in E. coli as a soluble protein. The obtained Gc protein showed good immunogenicity. Animal experiments showed that the rabbit serum antibody titer reached 1:512 000 after three immunizations. Conclusion In this study, SFTSV envelope glycoprotein Gc is successfully expressed in prokaryotic cells, which paves the way for subsequent studies of SFTSV envelope proteins.
YAO Wen-wu, YAN Hao, LOU Xiu-yu, PAN Jun-hang, SUN Yi, MAO Hai-yan, ZHANG Yan-jun
. Prokaryotic expression and immunogenicity of severe fever with thrombocytopenia syndrome virus glycoprotein Gc[J]. Chinese Journal of Vector Biology and Control, 2019
, 30(4)
: 383
-386
.
DOI: 10.11853/j.issn.1003.8280.2019.04.006
[1] Yu XJ,Liang MF,Zhang SY,et al. Fever with thrombocytopenia associated with a novel bunyavirus in China[J]. N Engl J Med,2011,364(16):1523-1532. DOI:10.1056/NEJMoa1010095.
[2] Liu Q,He B,Huang SY,et al. Severe fever with thrombocytopenia syndrome,an emerging tick-borne zoonosis[J]. Lancet Infect Dis,2014,14(8):763-772. DOI:10.1016/s1473-3099(14)70718-2.
[3] 施泽纶,乾虎,马世良,等. 新型布尼亚病毒病原学和诊断学进展[J]. 新发传染病电子杂志,2018,3(3):140-142.
[4] 吕沐天,孙颖,刘沛,等. 发热伴血小板减少综合征布尼亚病毒研究进展[J]. 微生物学杂志,2013,33(2):86-88.
[5] Takahashi T,Maeda K,Suzuki T,et al. The first identification and retrospective study of severe fever with thrombocytopenia syndrome in Japan[J]. J Infect Dis,2014,209(6):816-827. DOI:10.1093/infdis/jit603.
[6] Shin J,Kwon D,Youn SK,et al. Characteristics and factors associated with death among patients hospitalized for severe fever with thrombocytopenia syndrome,South Korea,2013[J]. Emerg Infect Dis,2015,21(10):1704-1710. DOI:10.3201/eid2110.141928.
[7] McMullan LK,Folk SM,Kelly AJ,et al. A new phlebovirus associated with severe febrile illness in Missouri[J]. N Engl J Med,2012,367(9):834-841. DOI:10.1056/NEJMoa1203378.
[8] Arikawa J,Yao JS,Yoshimatsu K,et al. Protective role of antigenic sites on the envelope protein of Hantaan virus defined by monoclonal antibodies[J]. Arch Virol,1992,126(1/4):271-281. DOI:10.1007/BF01309700.
[9] Överby AK,Pettersson RF,Grünewald K,et al. Insights into bunyavirus architecture from electron cryotomography of Uukuniemi virus[J]. Proc Natl Acad Sci USA,2008,105(7):2375-2379. DOI:10.1073/pnas.0708738105.
[10] 张文帅,迟莹,张黎,等. 发热伴血小板减少综合征布尼亚病毒Gn和Gc蛋白的分段表达[J]. 江苏预防医学,2013,24(1):4-6.DOI:10.3969/j.issn.1006-9070.2013.01.002.
[11] 徐菱遥,韩亚萍,周宜庆,等. SFTSV糖蛋白Gn重组质粒的构建及其体液免疫原性研究[J]. 南京医科大学学报:自然科学版,2016,36(5):554-558. DOI:10.7655/NYDXBNS20160509.
[12] 陈蕾,戴新宪,郝春生,等. 发热伴血小板减少综合征布尼亚病毒糖蛋白抗原(Gn)定量检测方法的建立及验证[J]. 中国生物制品学杂志,2017,30(10):1066-1072. DOI:10.13200/j.cnki.cjb.001747.
