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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (05): 96-102    DOI: Q789
    
Potential Industrial Production of Foreign Protein in Silkworm Using the Collusion Bodies of Recombinant Bombyx mori(silkworm) Multiple Nucleopolyhedrovirus Rescuesd by Polyhedrin Expressing Stable Cell Line
WU Kun1,2,FENG Juan1,2,YAO Lun-guang1,CHU Su-xia1,3,KAN Yun-chao1
1.ChinaUK, Nan Yang Normal UniversityRothamsted Research Joint Laboratory of Insect Biology, Henan Provincial Key Laboratory of Funiu Mountain Insect Biology, Nanyang Normal University, Nanyang 473061,China
2.Life Science College of Henan agriculture University, Zhengzhou 450002, China
3.Life Science College of Henan Normal University, Xinxiang 453007,China
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Abstract  

The transposon donor vector harboring both polyhedrin gene droved by BmA3 promoter and zeocin resistant gene controlled by IE1 promoter from OpNPV co-transefeced with a lepidoptera transposition helper plasmid pie2piggBac into BmN cells. A polyhedrin expressing stable cell line was constructed by screening with the 200μg/ml zeocin selection medium for one month. Occlusion bodies (OB) were rescued successfully in the polyhedrin expressing BmN cells infected with recombinant BmBac-gfp budded virus without polyhedrin gene. The amount of rescued OB produced in polyhedrin expressing cells is only 8% of wild type BmNPV OB produced in normal cells. The silkworm larvae were infected efficiently when they were feed by the rescued OB sprayed mulberry leaves. Foreign GFP was expressed efficiently and no OB was observed in the hemolymph of silkworm larvae. This rescued OB oral infection method is benefit to promote the industrial progress of silkworm bioreactor by eliminating the tedious injection budded virus solution by hand.

Key words： Rescuing cell line      Piggybac transposon      Occlusion body      Silkworm bioreactor     
Received: 31 December 2009      Published: 25 May 2010
Corresponding Authors: lunguang Yao     E-mail: lunguangyao@163.com
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TUN Kun
FENG Juan
TAO Lun-An
CHU Su-Xia
HAN Yun-Chao
Cite this article:

TUN Kun, FENG Juan, TAO Lun-An, CHU Su-Xia, HAN Yun-Chao. Potential Industrial Production of Foreign Protein in Silkworm Using the Collusion Bodies of Recombinant Bombyx mori(silkworm) Multiple Nucleopolyhedrovirus Rescuesd by Polyhedrin Expressing Stable Cell Line. China Biotechnology, 2010, 30(05): 96-102.

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https://manu60.magtech.com.cn/biotech/Q789     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I05/96

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