Gene transduction in mammalian cells using Bombyx mori nucleopolyhedrovirus assisted by glycoprotein 64 of Autographa californica multiple nucleopolyhedrovirus

被引:12
|
作者
Kato, Tatsuya [1 ,2 ,3 ]
Sugioka, Saki
Itagaki, Kohei
Park, Enoch Y. [1 ,2 ,3 ]
机构
[1] Shizuoka Univ, Res Inst Green Sci & Technol, Green Chem Res Div, Biotechnol Lab,Suruga Ku, 836 Ohya, Shizuoka 4228529, Japan
[2] Shizuoka Univ, Fac Agr, Dept Appl Biol Chem, Biotechnol Lab,Suruga Ku, Shizuoka 4228529, Japan
[3] Shizuoka Univ, Grad Sch Sci & Technol, Dept Biosci, Biotechnol Lab,Suruga Ku, Shizuoka 4228529, Japan
来源
SCIENTIFIC REPORTS | 2016年 / 6卷
关键词
ENVELOPE GLYCOPROTEIN; PROTEIN-PRODUCTION; FUSION PROTEIN; BACULOVIRUS; EXPRESSION; VIRUS; GP64; SILKWORM; SEQUENCE; LARVAE;
D O I
10.1038/srep32283
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Autographa californica multiple nucleopolyhedrovirus (AcMNPV), an alphabaculovirus, has been widely utilized for protein expression in not only insect cells but also mammalian cells. AcMNPV is closely related to Bombyx mori nucleopolyhedrovirus (BmNPV), and nucleotide sequences of AcMNPV genes have high similarity with those of BmNPV. However, the transduction of BmNPV into mammalian cells has not been reported. In this study, we constructed a recombinant BmNPV (BmNPV Delta bgp/AcGP64/EGFP) whose surface 64 kDa glycoprotein (BmGP64) was substituted with that from AcMNPV (AcGP64). BmNPV Delta bgp/AcGP64/EGFP also carried an EGFP gene under the control of the CMV promoter. BmNPV Delta bgp/AcGP64/EGFP successfully transduced HEK293T cells. In comparison, a control construct (BmNPV Delta bgp/BmGP64/EGFP) which possessed BmGP64 instead of AcGP64 did not express EGFP in HEK293T cells. The transduction efficiency of BmNPV Delta bgp/AcGP64/EGFP was lower than that of an AcMNPV based-BacMam GFP transduction control. This result indicates that AcGP64 facilitates BmNPV transduction into HEK293T cells. BmNPV can be prepared easily on a large scale because BmNPV can infect silkworm larvae without any special equipment, even though specific diet is needed for silkworm rearing. BmNPV gene transduction into mammalian cells can potentially be applied easily for gene delivery into mammalian cells.
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页数:9
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