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鸡法氏囊病毒重组多抗原表位串联肽的制备及其免疫反应
蔡梅红,谭仁可,韦瑶,段玉清,王亮,马海乐
0
(江苏大学食品与生物工程学院,镇江 212013)
摘要:
合成含有鸡法氏囊病毒抗原表位核酸序列的4条引物,利用SOE-PCR (重叠延伸PCR法)方法克隆得到含鸡法氏囊病毒多抗原表位串联肽的核酸序列。通过EcoR I和Sal I 两个酶切位点使该核酸序列插入原核表达载体(pET32a)。SDS-PAGE 实验结果表明鸡法氏囊病毒重组多抗原表位串联肽在大肠杆菌中的表达量为20%左右。Western blotting试验和免疫琼脂扩散沉淀试验(AGP)的结果均表明鸡法氏囊病毒重组多抗原表位串联肽具有明显的抗原性。
关键词:  法氏囊病毒  表位肽  抗原性
DOI:10.13610/j.cnki.1672-352x.20140620.004
投稿时间:2013-11-28
基金项目:国家自然科学基金(31201456), 江苏省博士后基金(1202042C), 江苏大学高级人才启动基金(12JDG099)和江苏省大学生实践创新训练计划(201310299076X)共同资助。
Development of recombinant multiple epitope peptide?of infectious bursal disease virus and its immune response
CAI Meihong,TAN Renke,WEI Yao,DUAN Yuqing,WANG Liang,Ma Haile
(School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013)
Abstract:
Four primers were synthesized to amplify nucleic acid sequences (a tandem repeat consisting of three IBDV multiple epitopes DNA sequence) by SOE PCR (splicing overlap extension PCR method). The nucleic acid sequence digested by EcoR I and Sal I was inserted into the expression vector pET32a. SDS-PAGE results showed that the recombinant multiple epitopes peptide (tandem peptide?of three IBDV epitopes) comprised about 20 % of the E.coli total protein. Agarose diffusion assay suggested that recombinant multiple epitopes peptide was demonstrated to have good antigenicity.
Key words:  infectious bursal disease virus  epitope peptide  antigenicity

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