Construction of a highly efficient display system for baculovirus and its application on multigene co-display

被引:14
|
作者
Zheng, Hao [1 ,2 ]
Wang, Xiong [1 ,2 ]
Ren, Feifei [1 ,2 ]
Zou, Shenglong [3 ]
Feng, Min [1 ,2 ]
Xu, Liangliang [1 ,2 ]
Yao, Lunguang [4 ,5 ]
Sun, Jingchen [1 ,2 ]
机构
[1] South China Agr Univ, Guangdong Prov Key Lab Agroanim Genom & Mol Breed, Coll Anim Sci, Guangzhou 510642, Guangdong, Peoples R China
[2] South China Agr Univ, Subtrop Sericulture & Mulberry Resources Protect, Coll Anim Sci, Guangzhou 510642, Guangdong, Peoples R China
[3] Guangzhou Cynosure Biotechnol Co Ltd, Guangzhou 510080, Guangdong, Peoples R China
[4] Nanyang Normal Univ, China UK NYNU RRes Joint Lab Insect Biol, Collaborat Innovat Ctr Water Secur Water Source R, Nanyang 473061, Henan, Peoples R China
[5] Nanyang Normal Univ, Henan Key Lab Ecol Secur Water Source Reg Midland, Collaborat Innovat Ctr Water Secur Water Source R, Nanyang 473061, Henan, Peoples R China
基金
中国国家自然科学基金;
关键词
Insect virus; Baculovirus multigene display system; Gene expression; High efficiency; NUCLEAR-POLYHEDROSIS-VIRUS; SWINE-FEVER VIRUS; SURFACE DISPLAY; ENVELOPE GLYCOPROTEIN; MAMMALIAN-CELLS; NONTRANSLATED REGION; INTERNAL INITIATION; ESCHERICHIA-COLI; FOREIGN PROTEINS; GENE-TRANSFER;
D O I
10.1007/s00438-018-1459-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The classical baculovirus display system (BDS) has often recruited fields including gene delivery, gene therapy, and the genetic engineering of vaccines, as it is capable of presenting foreign polypeptides on the membranes of recombinant baculovirus through a transmembrane protein. However, classical BDS's high cost, complicated operation, low display efficiency and its inability to simultaneously display multiple gene products impede its practicality. In this study, we present a novel and highly efficient display system based on ires-dependent gp64 for rescuing gp64-null Bacmid of baculovirus construction without affecting the viral replication cycle, which we name the baculovirus multigene display system (BMDS). Laser scanning confocal microscopy demonstrated that eGFP, eYFP, and mCherry were translocated on the membrane of Spodoptera frugiperda 9 cell successfully as expected. Western blot analysis further confirmed the presence of the fluorescent proteins on the budded, mature viral particles. The results showed the display efficiency of target gene on cell surface is fourfold that of classical BDS. In addition, a recombinant baculovirus displaying three kinds of fluorescent proteins simultaneously was constructed, thereby demonstrating the effectiveness of BMDS as a co-display system.
引用
收藏
页码:1265 / 1277
页数:13
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