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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2015, Vol. 35 Issue (8): 9-15    DOI: 10.13523/j.cb.20150802
研究报告     
轮状病毒VP4抗原表位在VP6载体蛋白同一位点表达比较研究
夏文跃1, 王晶1, 赵冰心1, 潘小霞2, 文喻玲1, 陈元鼎1
1. 中国医学科学研究院&北京协和医学院医学生物学研究所 云南省重大传染病疫苗研发重点实验室 昆明 650118;
2. 云南民族大学化学与生物技术学院 民族药资源化学国家民委-教育部重点实验室 昆明 650118
Comparative Analysis of Two Rotavirus VP4 Epitopes Inserted on the Same Site of VP6 Vector Protein
XIA Wen-yue1, WANG Jing1, ZHAO Bing-xin1, PAN Xiao-xia2, WEN Yu-ling1, CHEN Yuan-ding1
1. Chinese Academy of Medical Sciences & Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases, Institute of Medical Biology, Kunming 650118, China;
2. Key Laboratory of Chemistry in Ethnic Medicinal Resources, State Ethnic Affairs Commission and Ministry of Education, Yunnan Minzu University, Kunming 650500, China
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摘要：

目的: 评价轮状病毒(RV)VP4两个抗原表位插入VP6载体蛋白同一位点所表达的重组嵌合蛋白免疫学性质及在研制嵌合蛋白疫苗中的意义。方法: 采用分子克隆和基因重组技术将RV VP4的两个抗原表位插入到VP6载体蛋白同一位点上,构建重组抗原表达质粒,表达携带不同抗原表位的重组嵌合蛋白,用Western blot 和中和试验分析重组嵌合蛋白的抗原反应性和免疫原性。结果: 成功构建了两个嵌合蛋白表达质粒,并在大肠杆菌中高效表达;表达的嵌合蛋白可与相应抗体特异性反应;可诱导豚鼠产生特异性血清抗体;抗嵌合蛋白血清抗体可特异性识别载体蛋白VP6F,Wa株病毒的VP6和VP4蛋白,可中和Wa株病毒在MA104细胞上的感染性;结果表明,所构建和表达的两个以VP6为载体的VP4抗原表位嵌合蛋白具有较高抗原反应性和免疫原性;嵌合蛋白携带的VP4抗原表位具有增强载体蛋白免疫原性作用;为研制新型RV重组蛋白疫苗的奠定了较好的基础。
关键词： 轮状病毒;  抗原表位;  重组嵌合蛋白载体;  重组嵌合蛋白;  免疫原性    
Abstract:

Objective: To evaluate immunoreactivity and value of chimeric proteins that carry VP4 epitopes on the same insertion site using rotavirus VP6 as a vector. Methods: Two plasmids that carry genes of chimeric proteins carrying VP4 epitopes on the same insertion site using rotavirus VP6 as a vector were constructed by gene cloning and recombination; the chimeric proteins were expressed in E. coli cells, detected with anti-VP6 antibodies, as antigens inoculated in guinea pigs; and neutralization activities of antibodies in sera from chimeric proteins inoculated guinea pigs were determined. Results: the expressed chimeric proteins could react with antibodies against VP6 of rotavirus, could elicit production of antibodies in inoculated guinea pigs that reacted with vector protein VP6F, VP6 and VP4 of strain Wa virus, neutralized infectivities of Wa virus in MA104 cell. The results showed that chimeric proteins carrying different VP4 epitopes on the same insertion site retained its original backbone structure stability and had good immunoreactivity; results obtained are valuable for development of novel rotavirus vaccines.
Key words: Rotavirus    Epitopes    Epitope presenting vector    Recombinant chimeric protein    Immunoractivity
收稿日期: 2015-04-15 出版日期: 2015-08-25
ZTFLH:  Q789  
基金资助:

云南省自然科学基金资助项目(2013FZ130,2012FD039)
通讯作者: 陈元鼎     E-mail: chenyd@imbcams.com.cn
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引用本文:

夏文跃, 王晶, 赵冰心, 潘小霞, 文喻玲, 陈元鼎. 轮状病毒VP4抗原表位在VP6载体蛋白同一位点表达比较研究[J]. 中国生物工程杂志, 2015, 35(8): 9-15.

XIA Wen-yue, WANG Jing, ZHAO Bing-xin, PAN Xiao-xia, WEN Yu-ling, CHEN Yuan-ding . Comparative Analysis of Two Rotavirus VP4 Epitopes Inserted on the Same Site of VP6 Vector Protein. China Biotechnology, 2015, 35(8): 9-15.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150802    或    https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I8/9


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