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

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.

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 ultrapurification,nucleic acid electrophoresis,transmission electron microscope,and the liposome nanoparticles of50-100 nm were formed by liposome coating.After quantification,the gene nanoparticles were transfected into293T 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.