摘要
目的构建含有我国登革2型/4型病毒株嵌合E基因片段的真核表达质粒,观察重组质粒DNA的免疫原性,为登革多价疫苗的研究提供依据。方法首先将包含我国登革2型病毒43株E蛋白I/II抗原区和4型病毒B5株E蛋白III抗原区的嵌合E基因片段克隆至真核表达载体pcDNA3.1中,通过测序确定嵌合基因序列的正确性。然后将重组质粒以肌肉注射途径,免疫Balb/C小鼠。通过间接免疫荧光法对采集的鼠血清中的病毒特异抗体进行检测。结果构建的含有我国登革2型/4型病毒株嵌合E基因的真核表达质粒pcDNA-D2/4,经序列测定表明,导入的嵌合E基因片段的序列是正确的。将重组质粒DNA免疫小鼠,在初次免疫后的第3周,可同时检测到针对登革2型和4型病毒的特异荧光。结论所构建的含有不同血清型的我国登革病毒株嵌合E基因片段的真核重组质粒,可诱导小鼠同时产生针对两个血清型病毒的特异抗体,该研究为新型登革多价疫苗的研制提供了依据。
Aim To construct the eucaryotic recombi nant plasmid con-taining the chimaeric E gene from the Chinese dengue type 2and 4viruses,analyze the immunogenicit y of the recombinant plasmid DNA,and contribute to the study on multiv alent dengue vaccines.Methods The chimaeric E gene fragment contai ns the domains I /II of D2-43strain and domain III of D4-B5strain,and was cloned into the eu-caryotic plasmid pcDNA3.1.The sequ ence correctness of the chimaeric gene was affirmed by nucleotide sequencing.Then,Balb /C mice were immunized by the recombinant plasmi d with i.m ,and the specific an-tibodies to dengue viruses in the pooled sera were detected by IFA.Results The sequences of the chimaeric E gene contained in the recom-binant plasmid,named pcDNA -D2/4,w ere determined and showed to be exactly the same as the correspond ing parts of D2-43and D4-B5.Three weeks after the first immuniza tion,specific fluorescence was de-tected to both D2-43and D4-B5strain s.Conclusion By immunizing mice with recombinant eucaryotic ex pression plasmid containing the chi -maeric E gene from D2-43and D4-B5,sp ecific antibodies to both viruses were induced simultaneously,which conduces to the study on multivalent dengue vaccines.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第5期412-414,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助
No.39770036
