| 摘要: |
| 【目的】开发一种针对低密度脂蛋白成分ApoB100的新型重组酵母疫苗，为动脉粥样硬化(Atherosclerosis，AS)的防治提供一种思路。【方法】利用PCR得到包含编码ApoB100的 3 136-3 155位氨基酸残基的目的片段，将其克隆到酵母表达载体JMB88-HA-OVA-MCS上，构建JMB88-HA-OVA-hApoB载体，用硫酸铜诱导表达后获得HA-OVA-hApoB融合蛋白。采用口服的方法对昆明小白鼠免疫成功表达HA-OVA-hApoB蛋白的酵母细胞12周，每周1次，每次1×108个酵母细胞，免疫结束后4周采血，用Western blot检测小鼠是否产生特异性抗体。【结果】PCR扩增获得了859 bp的人ApoB100基因。成功构建了JMB88-HA-OVA-hApoB载体。口服免疫12周后，HA-OVA-hApoB免疫组小鼠的血清中含有特异性ApoB100抗体。【结论】灌服表达人ApoB100肽段融合蛋白的重组酿酒酵母能诱导小鼠的免疫反应，使其产生特异性抗体。 |
| 关键词: 昆明小白鼠 载脂蛋白 B 重组酵母疫苗 口服免疫 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目 (31172186) |
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| Induction of mice specific antibody by oral administration of whole recombinant yeast vaccine (ApoB100) |
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MA Li-xia,ZHANG Ting-ting,ZHANG You-you,et al
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| Abstract: |
| 【Objective】The objective of this study was to develop a novel recombinant yeast vaccine which targets the low-density lipoprotein apolipoprotein B-100 (ApoB100) to induce a protective immune response,and could improve the prevention and cure of atherosclerosis (AS).【Method】PCR was used to amplify a target fragment from human genomecontainng apoB-100,and clone it to express vetor JMB88-HA-OVA-MCS to construct JMB88-HA-OVA-hApoB vector.HA-OVA-hApoB fusion protein was obtained after induction using copper sulfate.Oral delivery of 1×108 recombinant yeasts with HA-OVA-hApoB protein to each mouse was conducted once a week for 12 weeks.Serum samples of immunized mice were collected 4 weeks after the immunization and analyzed using Western bolt method to check if ApoB100-specific antibody presented.【Result】A 859 bp human ApoB100 fragment was obtained by PCR.JMB88-HA-OVA-hApoB vector was established successfully.Oral administration of apoB-100-specific recombinant yeast vaccine for 12 weeks had a specific immune response to ApoB100 peptide.【Conclusion】Oral administration of recombinant yeast with ApoB100 fusion protein induced mice to create specific antibody. |
| Key words: mice apolipoprotein B recombinant yeast vaccine oral administration |
