Position: Home > Articles > Construction and Identification of Cell Penetrating Peptides VP22 Gene Fused with Enhanced Green Fluorescent Protein and Three Antigens of Toxoplasma gondii
China Animal Husbandry & Veterinary Medicine
2016,43
(11)
2859-2865
VP22与EGFP、3种弓形虫抗原融合表达重组质粒的构建与鉴定
作 者：
王娜;刘彦双;刘宁;金超;张喜路;金月;薛书江
单 位：
龙井市畜牧业管理局;龙井市畜牧站;延边大学农学院;梅河口市畜牧总站;大连市金州新区动物卫生监督所
关键词：
细胞渗透肽VP22;弓形虫;真核表达
摘 要：
为了探索新型弓形虫疫苗的传递系统,本试验分别构建了细胞渗透肽VP22与增强型绿色荧光蛋白(EGFP)融合表达的重组质粒(pcDNA-VP22-EGFP),以及细胞渗透肽VP22与3种弓形虫抗原融合表达的重组质粒(pcDNA-VP22-SAG1、pcDNA-VP22-GRA4、pcDNA-VP22-AMA1)。经PCR鉴定、酶切鉴定和序列测定后,将重组质粒pcDNA-VP22-EGFP转染COS7细胞,通过荧光显微镜和RT-PCR检测,验证VP22-EGFP基因在COS7细胞中的表达情况。PCR鉴定、酶切鉴定和序列测定结果显示所有重组质粒均构建正确。转染72h后的细胞,在荧光显微镜下成功观察到绿色荧光。RT-PCR扩增得到了大小为1 449bp的目的条带,与预期结果一致。结果表明,重组质粒构建成功。
译 名：
Construction and Identification of Cell Penetrating Peptides VP22 Gene Fused with Enhanced Green Fluorescent Protein and Three Antigens of Toxoplasma gondii
作 者：
WANG Na;LIU Yan-shuang;LIU Ning;JIN Chao;ZHANG Xi-lu;JIN Yue;XUE Shu-jiang;Agricultural College,Yanbian University;Longjing Animal Husbandry Station;Longjing Livestock Authority;Meihekou Animal Husbandry Station;Dalian Jinzhou New Area District Animal Health Supervision;
关键词：
cell penetrating peptides VP22;;Toxoplasma gondii;;eukaryotic expression
摘 要：
To study a novel vaccine of Toxoplasma gondii,the recombinant plasmids that cell penetrating peptides VP22 gene fused with one enhanced green fluorescent protein(EGFP)(pcDNAVP22-EGFP)and three antigens of T.gondii(pcDNA-VP22-SAG1,pcDNA-VP22-GRA4 and pcDNA-VP22-AMA1)were constructed,respectively.The recombinant plasmids were identified by PCR,restriction endonuclease digestion and DNA sequencing.The pcDNA-VP22-EGFP plasmid was transfected into COS7 cells,and the expression of VP22-EGFP in COS7 cells was confirmed by fluorescence microscope and RT-PCR.The results showed that all recombinant plasmid were constructed correctly.Green fluorescence was observed successfully under the fluorescence microscope in transfected cells after 72 h.The gene fragment of 1 449 bp was obtained by RTPCR.The results indicated that the recombinant plasmids were constructed successfully.
导出引用
计量
文章访问数: 7
HTML全文浏览量: 0
PDF下载量: 0
所属期刊
