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禽网状内皮组织增生症病毒p30-gp90串联表达蛋白及其免疫原性 被引量:2

Immunogenicity of co-expressed p30-gp90 against Reticuloendotheliosis virus
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摘要 【目的】研究禽网状内皮组织增生症病毒(Reticuloendotheliosis Virus,REV)群特异性抗原P30与囊膜糖蛋白gp90体外共表达蛋白的免疫原性,为研发新型REV抗体诊断试剂盒提供基础。【方法】根据REV脾脏坏死病毒(spleen necrosis virus,SNV)株的前病毒基因组cDNA序列,设计合成2对引物,以pPB101质粒为模板,分别扩增REV p30基因和gp90基因片段。将PCR产物依次克隆入表达载体pET-28a(+)中,通过酶切鉴定和测序分析,筛选阳性重组克隆pET-p30-gp90。重组菌经异丙基硫代D-半乳糖苷(IPTG)诱导后,通过SDS-PAGE电泳分析表达情况,Western blot检测表达蛋白与特异性血清之间的反应性。制备表达蛋白的抗血清,以该抗血清与REV感染的鸡胚成纤维细胞(CEF)进行间接免疫荧光实验(IFA),验证表达蛋白的免疫原性。【结果】经SDS-PAGE电泳后能观察到预期大小的表达条带,Western blot结果显示,重组蛋白能与REV抗血清反应。将表达产物纯化后免疫Balb/c小鼠,制备p30-gp90抗血清,该抗血清与REV感染CEF在IFA中呈现特异性荧光反应。【结论】体外串联表达REV p30-gp90蛋白,表达蛋白具有良好的免疫原性。 [Objective] To study the immunogenicity of co-expression of p30,one of a group specific antigens,and glycoprotein gp90 genes of Reticuloendotheliosis virus (REV).[Methods]p30 and gp90 genes were amplified with the template of plasmid pPB101 containing the whole sequence of spleen necrosis virus(SNV),and cloned into pET-28a(+) vector.The positive clone pET-p30-gp90 was identified by enzyme analysis and sequencing.The co-expressed protein p30-gp90 was confirmed by SDS-PAGE and Western blot analysis. After quantitation,the purified protein p30-gp90 was immunized to Balb/c mice three times to prepare the p30-gp90 specific anti-serum.Afterwards,we detected the REV infected Chick Embryo Fibroblasts (CEF) by Immunofluorescence assay (IFA) with the prepared anti-serum.[Results]The p30-gp90 was co-expressed efficiently by SDS-PAGE and Western blot analysis.The anti-serum of p30-gp90 reacts with REV infected CEF in IFA.[Conclusion]The expressed protein of p30-gp90 keeps good immunogenicity.
出处 《微生物学报》 CAS CSCD 北大核心 2014年第3期352-358,共7页 Acta Microbiologica Sinica
基金 重大动物疫病新型疫苗的关键技术研究与产业化(2011A090200117)~~
关键词 禽网状内皮组织增生症病毒 P30 gp90 免疫原性 Reticuloendotheliosis virus, p30, gp90, immunogenicity
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  • 1崔治中 孙怀昌 等.禽白血病及禽网状内皮增生病感染情况的调查[J].中国畜禽传染病,1987,(1):37-38.
  • 2Kang CY, Temin HM. Lack of sequence homology among RNAs of avian leucosis sarcoma viruses reticuloendotheliosis viruses, and chicken endogenous RNA directed DNA polymerase activity. Journal of Virology, 1973, 12:1314-13241.
  • 3李广兴,任晓峰,杨贵君,刘忠贵.网状内皮组织增殖病病毒感染SPF雏鸡后病毒在各器官内的分布[J].中国兽医科技,1999,29(11):6-8. 被引量:11
  • 4Witter RL. Reticuloendotheliosis.// Calnek BW. Disease of Poultry. 10'h eds. Ames, USA: Iowa State University Press, 1997. 467-484.
  • 5胡北侠,黄艳艳,路希山,孟斌,张秀美.肉种禽网状内皮增生症、鸡传染性贫血和禽白血病血清学调查[J].广东畜牧兽医科技,2009,34(1):25-27. 被引量:30
  • 6Lovinger GG, Mark G, Todaro GJ, Schochetman G. 5 '-terminal nucleotide noncoding sequence sof retroviruses : relatedness of two old world primate type C viruses and avian spleen necrosis virus. Journal of Virology, 1981, 39:238-2451.
  • 7Rice NR, BonnerTI, Gilden RV. Nucleic acid homology between avian and mammalian type C viruses: relatedness of reticuloendotheliosis virus cDNA to cloned proviral DNA of the endogenous colobus virus CPC-11. Virology, 1981,114:286-2901.
  • 8Weaver TA, Talbot K J, Panganiban AT. Spleen necrosis virus gag polyprotein in necessary for particle assembly and release but not for proteolytic processing. Virology, 1990, 64: 2642-2652.
  • 9Jackson CA, Dunn SE, Smith DI, Gilchrist PT, Macqueen PA. Proventriculitis, " Nakanuke " and reticuloendotheliosis in chickens following vaccination with herpesvirus of turkeys. Journal of Veterinary, 1977, 53: 457- 458.
  • 10吉荣,崔治中,王锡乐,孙淑红.分子克隆化禽网状内皮组织增生症病毒传染性及其前病毒全基因组序列研究[J].病毒学报,2005,21(6):448-455. 被引量:17

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