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携带双串联卵清蛋白T细胞表位的嵌合兔出血症病毒衣壳蛋白自聚能力的研究

DOI: 10.7668/hbnxb.2014.01.011, PP. 55-61

Keywords: 兔出血症病毒,VP60,双串联OVAT细胞表位,病毒样颗粒

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Abstract:

以兔出血症病毒(RHDV)衣壳蛋白VP60为载体,构建VP60携带双串联OVAT细胞表位(Ovalbumin,卵清蛋白第257~264位氨基酸)的嵌合蛋白,研究外源基因对VP60蛋白表达及病毒样颗粒(VLPs)装配的影响,分析VP60作为载体对外源基因长度的耐受性。运用基因克隆和重组技术,通过2次SOE法(Splicingbyoverlapextension,重叠延伸剪接术),将双串联OVAT细胞表位(257~264位氨基酸)的基因序列取代VP60第302~309位氨基酸的基因序列,得到嵌合VP60基因。利用杆状病毒表达系统表达嵌合蛋白,命名为DC。经IFA、SDS-PAGE、WesternBlot、RT-PCR方法鉴定嵌合蛋白的表达,并通过电镜观察嵌合蛋白自聚为病毒样颗粒的能力。嵌合VP60蛋白在杆状病毒表达系统中得到高效表达,且可自聚形成病毒样颗粒。携带双串联外源表位的嵌合VP60蛋白得到有效表达,并可形成VLPs。该研究为嵌合VP60VLPs的形成、结构功能的研究奠定基础,同时扩展了VP60作为载体可携带的外源基因耐受性的研究资料。

References

[1]  3 Martin-Alonso J M,Casais R, Boga J A, et al. Processingof rbabbit hemorrhagic disease virus polyprotein[J]Journal of Virology,1996,70 (2):1261-1265
[2]  4 Barcena J,Morales M,Vazquez B,et al. Horizontal trans-missible protecaion against myxomatosis and rabbit hemor-rhagic disease by using a recombinant myxoma virus[J]Journal of Virology,2000,74:1114-1123
[3]  5 Farnos 0,Boue 0,Parra F, et al. High-level expressionand immunogenic properties of the recombinant rabbithemorrhagic disease virus VP60 capsid protein obtainedin Pichia pastoris[J]J Biotech,2005,117(3):215-224
[4]  6 Mao C,solis D J, Reiss B D,et al. Virus-based toolkitforthe direcaed synthesis of magnetic: and semiconducaingnanowires[J]. Science,2004 (303);213-217
[5]  7 Wang Q,Lin T, Tang L, et al. Icosahedral virus particaesas addressable nanoscale building blocks[J}. AngewChe-mint EdEng1,2002(41);459-462
[6]  17 Wang X, Xu F, Liu J, et al. Atomic: model of rabbit hem-orrhagic disease virus by cryo-elecaron microscopy and crystallography [J]. PLoS Pathogens,2013, 9:1003132
[7]  18 Chen Rong, Neill John D,Noel Jacqueline S, et al. Inter-and intragenus strucaural variations in caliciviruses andtheir funcaional implications[J]Journal of Virology,2004;78(12);6469-6479
[8]  1 蔡宝祥家畜传染病学第三版[M]北京:中国农业出版社,1998;299-301
[9]  2 Wirblich C,Thiel H J,Meyers G. Genetic: map of the cali-civirus rabbit hemorrhagic disease virus as deduced fromIn vitro translation studies [J]. Journal of Virology,1996,70(11);7974-7983
[10]  8 Jennings G T, Bachmann M F. The coming of age of virus-like particle vaccines[J]Biol Chem, 2008,389(5):521-536
[11]  9 Ludwig C,Wagner R. Virus-like particaes-universal mo-lec:ular tool boxes[J]CurrOpin Biotechnol, 2007,18(6);537-545
[12]  10 Barcena J, Verdaguer N,Roc:a R,et al. The coat protein of Rabbit hemorrhagic disease virus contains a molecular switch at the N-terminal region facing the inner surface of the capsid[J]. Virology,2004,322:118-134
[13]  11 Nagesha H S,Wang L F,Hyatt A D. Virus-like particaesof calicivirus as epitope carriers[J]. Archives of Virolo-gy,2000,144(12);2429-2439
[14]  12 Crisci E,Almanza H,Mena I, et al. Chimeric calici-virus-like particaes elicit protecaive anti-viral cytotoxicresponses with out adjuvant[J]Virology, 2009,387: 303-312
[15]  13 王芳,胡波,任雪枫,等兔出血症病毒衣壳蛋自在昆虫细胞中的表达及对家兔的免疫保护效果[J]畜牧兽医学报,2008,39(10);1382-1387
[16]  14 蔡少平,王芳,贾华敏,等兔出血症病毒胶体金免疫层析试纸条诊断方法的建立及初步应用[J]畜牧兽医学报,2012,43(11):1795-1801
[17]  15 杨廷亚,王芳,姜平,等应用噬菌体展示技术筛选兔出血症病毒抗原模拟表位[J]畜牧兽医学报,2012,43(8):1281-1286
[18]  16 Chac:kerian Bryc:e. Virus-like particaes:flexible platformsfor vaccine development[J]Expert Review of Vac-cines,2007,6;381-390

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