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鹅细小病毒VP3蛋白的原核表达及纯化

Prokaryotic expression and purification of VP3 protein of goose parvovirus
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摘要 目的原核表达并纯化鹅细小病毒(Goose parvovirus,GPV)VP3蛋白。方法将GPV VP3基因重组表达质粒pGEX-6P-VP3转化入大肠杆菌BL21(DE3)plysS内,IPTG诱导表达;用Sepharose 4B(Prepacked Glutathion)亲合层析柱纯化可溶性和不溶性包涵体蛋白,并进行SDS-PAGE、Western blot及Dot-ELISA鉴定。结果表达的GST/VP3融合蛋白相对分子质量为85 000,主要以不溶性包涵体形式存在;纯化的可溶性蛋白和包涵体蛋白含量分别为2.20和3.30 mg/ml;纯化的GST/VP3融合蛋白能被鹅抗GPV多克隆抗体特异性识别。结论原核表达并纯化了GPV VP3蛋白,为进一步研究其功能和免疫学特性及研制基因工程亚单位疫苗和新型抗原制剂奠定了基础。 Objective To express the VP3 protein of goose parvovirus (GPV) in prokaryotic cells and purify the expressed product. Methods Recombinant plasmid pGEX-VP3 was transformed into E. coli BL21 (DE3) plysS and induced with IPTG. The obtained soluble protein and inclusion body were purified by Sepharose 4B (Prepacked Glutathione) affinity chromatography, and i- dentified by SDS-PAGE, Western blot and Dot-ELISA. Results The expression fusion protein GST/VP3, with a relative molecular mass of 85 000, mainly existed in a form of insoluble inclusion body. The contents of purified soluble protein and inclusion body were 2. 20 and 3. 30 nag/ml respectively. Purified fusion protein GST/VP3 was specifically recognized by goose anti-GPV polyclonal antibody. Conclusion VP3 protein of GPV was expressed in prokaryotic cells and purified, which laid a foundation of further study on its function and immunological characters and development of recombinant subunit vaccine and novel antigen preparation.
出处 《中国生物制品学杂志》 CAS CSCD 2012年第10期1307-1309,1314,共4页 Chinese Journal of Biologicals
关键词 鹅细小病毒 VP3蛋白 原核细胞 基因表达 纯化 Goose parvovirus (GPV) VP3 protein Prokaryotic cells Gene expression Purification
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  • 1Ji J, Xie QM, Chen CY, et al. Molecular detection of Muscovy duck parvovirus by loop-mediated isothermal amplication assay [J]. Pouh Sci, 2010, 89(3): 477-483.
  • 2邓尚龙,黄仁美,王生育,颜江华,江清银.鹅细小病毒VP3基因的克隆与表达[J].中国兽医杂志,2009,45(6):29-31. 被引量:2
  • 3Yu TF, Ma B, Gao MC, et al. Localization of liner B-cell epi- topes on goose parvovirus structural protein [J]. Vet Immunol Immunopathol, 2012,145 (1-2) : 522-526.
  • 4Ju HY, Wei N, Wang Q, et al. Goose parvovirus structural pro- tein expressed by recombinant baculoviruses self-assemble into virus-like particles with strong immunogenicity in goose [J]. Biochem Biophys Res Commun, 2011,409 ( 1 ) : 131-136.
  • 5布日额,王君伟,吴金花.检测小鹅瘟感染抗体的Dot-ELISA方法研究[J].中国生物工程杂志,2009,29(9):68-70. 被引量:4
  • 6Zhang Y, Li YF, Liu M, et ol. Development and evaluation of a VP3-ELISA for the detection of goose and Muscovy duck par- vovirus antibodies [J]. J Virol Methods,2010,163 (2) : 405- 409.
  • 7Zhu LQ, Ding XY, Tao J, et al. Identification of target cells for goose parvovirus infection in the immune system organs [J]. Acta Virol, 2010, 54 (3): 211-215.

二级参考文献12

  • 1布日额,李宝臣,马波,王君伟,Ulrich Neumann.应用GPV VP3基因重组原核表达产物建立检测抗体的ELISA方法研究[J].畜牧兽医学报,2006,37(2):199-203. 被引量:16
  • 2Schettler C H. Isolation of a highly pathogenic virus from geese with hepatitis [J].Avian Dis,1971,159(2):323-325.
  • 3蔡宝祥.家畜传染病学[M].3版.北京:中国农业出版社,2000:283-285.
  • 4Derzy D, Dren C, Sze do M, etal. Viral disease of gosling: I solation, properties and antigenic pattern of the virus strains [J]. ActaVet Hung, 1970, 20: 194-228.
  • 5Gough R E, Spackman D, Collins M S. Isolation and characterization of a parvovirus from goslings[J].Vet Rec, 1981, 108:399-400.
  • 6Le Gall-Recule G, Jestin V, Changnaug P, et al. Expression of muscovy duck parvovirus capsid proteins(VP2 and VP3) in a baculovirus expression system : a demons tration of immunity induced by the recombinant proteins[J]. J Gen Virol, 1993,77: 2159-2163.
  • 7Zadori Z, Raymund S, Tibor R, et al. Analysis of the corn plete nucleotide sequence of goose and Muscovy duck parvovir us indicates common ancestral origin with adeno-associated vi rus 2[J].Viroligy,1995,212(2):562-573.
  • 8Takehara K, Nishio, Hayashi Y, et al. Anout break of goose parvovirus infection in japan. J Vet Med Sci, 1995, 57 (4) :777 - 779.
  • 9BW卡尔尼克.禽病学.第十版.高福,苏敬良,译.北京:中国农业出版社,1999.988-990.
  • 10布日额,王君伟,吴金花,贾伟星,马波,范铁力,李勐,Ulrich Neumann,李哲迁.GPV VP1-VP3非重叠序列重组原核表达多肽的应用研究[J].中国兽医杂志,2007,43(7):22-23. 被引量:2

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