周健, 左原源, 赵雨芊, 赖清润, 郑惠文, 赵鑫, 秦丽, 张兴龙, 施海晶, 刘龙丁, 李恒. 脂质体包被SARS-CoV-2 N基因的候选DNA疫苗转染人源细胞效率研究[J]. 云南大学学报(自然科学版), 2022, 44(5): 1062-1068. doi: 10.7540/j.ynu.20210652
引用本文: 周健, 左原源, 赵雨芊, 赖清润, 郑惠文, 赵鑫, 秦丽, 张兴龙, 施海晶, 刘龙丁, 李恒. 脂质体包被SARS-CoV-2 N基因的候选DNA疫苗转染人源细胞效率研究[J]. 云南大学学报(自然科学版), 2022, 44(5): 1062-1068. doi: 10.7540/j.ynu.20210652
ZHOU Jian, ZUO Yuan-yuan, ZHAO Yu-qian, LAI Qing-run, ZHENG Hui-wen, ZHAO Xin, QIN Li, ZHANG Xing-long, SHI Hai-jing, LIU Long-ding, LI Heng. Efficiency of candidate DNA vaccine into anthropogenic cells with SARS-CoV-2 N gene encapsulated by liposomes[J]. Journal of Yunnan University: Natural Sciences Edition, 2022, 44(5): 1062-1068. DOI: 10.7540/j.ynu.20210652
Citation: ZHOU Jian, ZUO Yuan-yuan, ZHAO Yu-qian, LAI Qing-run, ZHENG Hui-wen, ZHAO Xin, QIN Li, ZHANG Xing-long, SHI Hai-jing, LIU Long-ding, LI Heng. Efficiency of candidate DNA vaccine into anthropogenic cells with SARS-CoV-2 N gene encapsulated by liposomes[J]. Journal of Yunnan University: Natural Sciences Edition, 2022, 44(5): 1062-1068. DOI: 10.7540/j.ynu.20210652

脂质体包被SARS-CoV-2 N基因的候选DNA疫苗转染人源细胞效率研究

Efficiency of candidate DNA vaccine into anthropogenic cells with SARS-CoV-2 N gene encapsulated by liposomes

  • 摘要: 利用脂质体包被含有SARS-CoV-2核衣壳蛋白(nucleocapsid protein;N)基因的候选DNA疫苗,并检测其在人源细胞293T中的转染效率. 将SARS-COV-2 N基因克隆入真核表达载体pcDNA3.1,DNA测序及转染后的质谱测序证明能够表达正确蛋白. 利用脂质体包被pcDNA3.1-SARS-COV-2-N基因,并进行超离纯化、核酸电泳、透射电镜等检测发现形成50~100 nm的脂质体纳米颗粒,定量后将纳米颗粒转染293T细胞,并对比脂质体转染与FUGENE®HD试剂转染效果,表明制备的候选重组脂质体DNA疫苗能够形成稳定的纳米颗粒,并可在人源细胞293T中高效表达.

     

    Abstract: We coated the candidate DNA vaccine containing SARS-CoV-2 nucleocapsid gene with liposome and detected its transfection efficiency in human cells 293T. SARS-CoV-2 N gene was cloned into eukaryotic expression vector pcDNA3.1(+) and identified by DNA sequencing and mass spectrometry sequencing. Then, the pcDNA3.1(+)-SARS-CoV-2 N gene nanoparticles coated by liposomes were prepared and detected by ultra-purification, nucleic acid electrophoresis, transmission electron microscope, and the liposome nanoparticles of 50−100 nm were formed by liposome coating. After quantification, the gene nanoparticles were transfected into 293T cells according to gradient concentration, and the transfection effects between liposome transfection and FUGENE ®HD transfection were compared. The results validated that the candidate recombinant liposome DNA vaccine formed stable nanoparticles with high efficiency expression in human cells 293T.

     

/

返回文章
返回