一种基于植物瞬时表达研制新冠病毒重组蛋白疫苗的方法  

A method of developing SARS-CoV-2 recombinant protein vaccine based on plant transient expression

作　　者：王弘瑞 赵丽娟[1] 房明丽[1] 杨明[1] WANG Hongrui;ZHAO Lijuan;FANG Mingli;YANG Ming(Department of Molecular Biology,College of Basic Medicine Sciences,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学基础医学院分子生物学教研室,长春130021

出　　处：《中国免疫学杂志》2022年第20期2512-2516,共5页Chinese Journal of Immunology

基　　金：吉林大学白求恩计划科研项目(2018B16)。

摘　　要：目的:探究一种以植物烟草为瞬时表达载体快速且高效研制和表达SARS-CoV-2重组蛋白疫苗的方法。方法:培育植物烟草,当烟草生长到第6周时,叶片生长到接近手掌大小,可用于下一步的农杆菌渗透。构建包含SARS-CoV-2 S重组质粒的农杆菌。培养含目的基因的农杆菌,来获得农杆菌渗透液。选择待渗透的目标叶片,用针头轻轻在叶片上刺穿一个小孔,然后将制备好的农杆菌渗透液注入叶片,使其慢慢吸收渗透液。需要从渗透后的烟叶中提取和纯化SARS-CoV-2 S重组蛋白,并进行初步鉴定。对SARS-CoV-2 S重组蛋白进行纯化与鉴定,并对纯化的SARS-CoV-2 S重组蛋白进行SDS-PAGE和Western blot分析。结果:采用植物烟草成功地表达SARS-CoV-2 S重组蛋白,其分子量约为53.2 kD,Western blot结果显示His-tag抗体可以识别SARS-CoV-2 S重组蛋白。结论:基于植物的瞬时表达系统可作为SARS-CoV-2重组蛋白疫苗的快速研制方法。Objective:To explore a method for rapid and efficient development and expression of SARS-CoV-2 recombinant protein vaccine using plant tobacco as transient expression vector.Methods:The plant tobacco was bred so that when the tobacco grew to 6 week,the leaves grew to nearly palm size and could be used for agrobacterium infiltration in the next step.Agrobacterium tumefaciens containing SARS-CoV-2 S recombinant plasmid were construsted.Agrobacterium tumeto containing target gene was cultured to obtain Agrobacterium tumeto osmotic solution.The target leaf to be penetrated was selected,a small hole was gently pierced in the leaf with a needle,and the prepared Agrobacterium permeate was injected into the leaf to allow it to slowly absorb the permeate.The SARSCoV-2 S recombinant protein neededs to be extracted and purified from the penetrated tobacco leaves and preliminarily identified.The SARS-CoV-2 S recombinant protein was purified and identified.The purified SARS-CoV-2 S recombinant protein was analyzed by SDS-PAGE and Western blot.Results:The recombinant SARS-CoV-2 S protein was successfully expressed in plant tobacco,and its molecular weight was about 53.2 kD.Western blot results also showed that His-tag antibody could recognize the recombinant SARSCoV-2 S protein well.Conclusion:The plant-based transient expression system can be used as a rapid method for preparation of novel coronavirus recombinant protein vaccine.

关 键 词：COVID-19 植物生产疫苗 瞬时表达系统 本氏烟草 农杆菌渗透法 

分 类 号：R392[医药卫生—免疫学]