VP22与EGFP、3种弓形虫抗原融合表达重组质粒的构建与鉴定

doi:10.16431/j.cnki.1671-7236.2016.11.008

Abstract

Abstract:

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)(pcDNA-VP22-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 RT-PCR.The results indicated that the recombinant plasmids were constructed successfully.

Key words: cell penetrating peptides VP22; Toxoplasma gondii; eukaryotic expression

CLC Number:

WANG Na, LIU Yan-shuang, LIU Ning, JIN Chao, ZHANG Xi-lu, JIN Yue, XUE Shu-jiang. Construction and Identification of Cell Penetrating Peptides VP22 Gene Fused with Enhanced Green Fluorescent Protein and Three Antigens of Toxoplasma gondii[J]. , 2016, 43(11): 2859-2865.

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Construction and Identification of Cell Penetrating Peptides VP22 Gene Fused with Enhanced Green Fluorescent Protein and Three Antigens of Toxoplasma gondii

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