同济大学学报（医学版）2011，Vol.32Issue(6)：1-8,8.DOI:10.3969/j.issn1008-0392.2011.06.001
禽流感病毒H5N1亚型基因工程疫苗设计、表达制备及动物实验研究
Preparation of avian influenza virus H5N1 genetic engineering vaccine and animal study
刘学东 1包振民 2王志亮1
作者信息
- 1. 中国海洋大学海洋生物遗传学与种质工程实验室,山东青岛266003
- 2. 青岛市市立医院呼吸科,山东青岛266011
- 折叠
摘要
Abstract
Objective To design and prepare a genetic engineering vaccine against avian influenza virus H5N1. Methods The composite amino acid sequences plus epitope subunit of vaccine were designed based on the analysis of prevalent avian influenza H5N1 strains and bioinformatics. The amino acid sequence was obtained with E. Coli codon optimization technique; the vaccine gene was synthesized with the whole gene synthesis method and expressed with E. Coli expression system. Theprotein inclusion bodies were purified by renaturation chromatography after washing; the antigen specificity was measured by ELISA. The emulsified protein vaccine of avian influenza prepared was evaluated by immunization test in mice for immune effect. Results The target protein expressed in E. Coli accounted for approximately 30% of the total bacterial protein; the purity of protein was 95. 5% after purification and the concentration of soluble protein was up to 2.4 mg/ml. Immune tests proved that the vaccine effectively induced immune responses in mice. Conclusion The engineering vaccine against avian influenza virus H5N1 has been successfully prepared which can induce immune response in mice.关键词
禽流感/疫苗/原核表达/蛋白质纯化/蛋白质复性/H5N1/免疫效力/小鼠Key words
Avian influenza/vaccine/prokaryotic expression/protein purification/protein refolding/H5N1/immune effect/mouse分类
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刘学东,包振民,王志亮..禽流感病毒H5N1亚型基因工程疫苗设计、表达制备及动物实验研究[J].同济大学学报（医学版）,2011,32(6):1-8,8.
