Improvement of recombinant baculovirus infection efficiency to express manganese superoxide dismutase in silkworm larvae through dual promoters of Pph and Pp10.
The silkworm, Bombyx mori, has been used as an important bioreactor for the production of recombinant proteins through baculovirus expression system (BES). There are several problems which will probably be the bottleneck for practical and industrial utilization of silkworm bioreactor. Traditionally, the recombinant virus should infect the larvae through individual dorsal injection by a syringe. This is a time- and labor-consuming procedure. This drawback has become a bottleneck for practical and industrial utilization of baculovirus expression system in the silkworm bioreactor. In this paper, we constructed a dual expression baculovirus to express the renovated polyhedron and target manganese superoxide dismutase (SOD) gene under P10 and polyhedron promoters, respectively, through oral infection. The results showed that the direct injection of recombinant rBacmid/BmNPV/SOD DNA with cellfectin reagent infected the silkworm larvae partially. When next batches of larvae were fed orally with hemolymph, which was collected from first batch of injected and infected larvae, the obvious symptom of infection was found and high target SOD was expressed. These results imply it is feasible to express target genes through combination of recombinant bacmid DNA injection and oral feeding by a dual expression bacmid baculovirus.
Yue
Wan-fu
WF
Institute of Sericulture and Apiculture, Zhejiang University, Hangzhou, 310029, People's Republic of China.
Li
Xing-hua
XH
Wu
Wei-cheng
WC
Roy
Bhaskar
B
Li
Guang-li
GL
Liu
Jian-mei
JM
Wu
Xiao-feng
XF
Zhou
Ji-yong
JY
Zhang
Chuan-xi
CX
David
Wan Chi Cheong
WC
Miao
Yun-gen
YG
D000818:Animals; D012831:Bombyx / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006458:Hemolymph / Q000821:virology; D007814:Larva / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D000077183:Occlusion Body Matrix Proteins; D010714:Phosphatidylethanolamines; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics*
D004279:DNA, Viral; D000077183:Occlusion Body Matrix Proteins; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; C020888:1,2-dielaidoylphosphatidylethanolamine; D013482:Superoxide Dismutase