摘要
目的 利用毕赤酵母系统表达重组新型冠状病毒受体结构域蛋白RBD,以期作为新冠病毒重组蛋白候选疫苗。方法 选择新型冠状病毒的RBD(受体结合域)蛋白为靶点,通过基因工程技术将RBD基因序列克隆到毕赤氏酵母表达载体pPIC9K中。通过载体将外源基因整合到毕赤酵母染色体上,获得遗传性稳定重组子。毕赤氏酵母表达载体含有醇氧化酶基因的调控序列,加入甲醇进行诱导,通过前导信号肽引导外源蛋白的分泌表达。结果 成功将RBD基因序列克隆到表达载体中并整合到酵母染色体上,获得遗传性稳定重组子。能通过前导信号肽引导目的蛋白的分泌表达,表达产物免疫动物刺激产生高水平的血清IgG抗体,IgG抗体效价为1∶2.73×106。结论 重组新型冠状病毒受体结构域蛋白RBD的设计具有可行性,为重组蛋白候选疫苗在预防新型冠状病毒感染中具有潜在的应用价值。
Objective The Pichia pastoris system was used to express the recombinant 2019-nCoV receptor domain protein RBD to develop a 2019-nCoV recombinant protein candidate vaccine.MethodsWe selected the RBD(receptor binding domain)protein of the new coronavirus as the target,and cloned the RBD gene sequence into the expression vector pPIC9K of Pichia pastoris through genetic engineering technology.Results The RBD gene sequence was successfully cloned into an expression vector and integrated into the yeast chromosome to obtain genetically stable recombinants.It can guide the secretion and expression of the target protein through the leading signal peptide,and the expression product can stimulate the immunized animals to produce high levels of serum IgG antibodies,and the IgG antibody titer is 1:2.73 X10°.Conclusion Antigen design is feasible.The vaccine has potential application value in the prevention of 2019-nCoV infection,and the results of this study provide an effective strategy and method for the development of 2019-nCoV vaccine.
作者
张桐
杨松惠
姜人月
唐家凤
许智强
狄亚心
于潼
方效良
田明尧
金宁一
庄忻雨
朱光泽
ZHANG Tong;YANG Songhui;JIANG Renyue;TANG Jiafeng;XU Zhiqiang;DI Yaxin;YU Tong;FANG Xiaoliang;TIAN Mingyao;JIN Ningyi;ZHUANG Xinyu;ZHU Guangze(College of Veterinary Medicine,Jilin University,Changchun,Jilin 130062,China;Changchun Institute of Veterinary Medicine,Chinese Agricultural Sciences;Zhejiang Dongfang Biological Gene Products Co,Ltd.;Affiliated Hospital of Changchun University of Traditional Chinese Medicine)
出处
《中国病原生物学杂志》
CSCD
北大核心
2023年第11期1241-1244,1251,共5页
Journal of Pathogen Biology
基金
新型冠状病毒肺炎变异株重组蛋白疫苗研发项目(No.21ZGY30)。
