摘要
目的筛选和鉴定猴B病毒囊膜蛋白gB的特异性抗原表位,将其应用于B病毒的检测。方法利用蛋白序列比较和表位预测技术筛选猴B病毒囊膜蛋白gB的特异性抗原表位,经PCR扩增后原核表达,纯化,Western-blot鉴定融合蛋白,建立特异性表位的ELISA检测方法 ,并对其效果进行评估。结果琼脂糖凝胶电泳和测序结果显示出目的表位基因完全正确,并且重组蛋白经过SDS-PAGE、Western-blot鉴定,其相对分子质量约为27×103,与预期值相符。筛选出的gB-26肽表位检测结果与文献相符,特异性较好,敏感性稍低。结论建立了猴B病毒囊膜蛋白gB特异性抗原表位筛选和鉴定的实验方法 ,为进一步研制猴B病毒快速诊断试剂盒和猴B病毒亚单位疫苗奠定了基础。
Objective To select and identify B virus envelop protein gB specific antigen epitope and to apply to the detection of B virus. Methods Using protein sequence comparison and epitope prediction to select the gB specific antigen eitope. Then obtaining the fusion protein by prokaryon expression and Western-blot assessment,building the ELISA method to detect epitope and evaluating the effects. Results The gene sequence of epitopewas completely correct,the molecular weight of recombination protein identifying by Western-blot was about 27 × 10^3. The gB-26 epitide was better in specificity and lower in sensitivity. Conclusion Building up methods to selecet and identify specific antigen eitope.
出处
《中国比较医学杂志》
CAS
2010年第10期10-14,87,共6页
Chinese Journal of Comparative Medicine
基金
军事医学科学院科研创新基金资助
关键词
B病毒
囊膜蛋白gB
抗原表位
筛选
表达
B virus
Envelop protein gB
Antigen epitope
Selection
Identification
