Improvement of recombinant baculovirus infection efficiency to express manganese superoxide dismutase in silkworm larvae through dual promoters of Pph and Pp10

• Published in: Applied microbiology and biotechnology 2008-03, Vol.78 (4), p.651-657
• Main Authors: Yue, Wan-fu, Li, Xing-hua, Wu, Wei-cheng, Roy, Bhaskar, Li, Guang-li, Liu, Jian-mei, Wu, Xiao-feng, Zhou, Ji-yong, Zhang, Chuan-xi, David, Wan Chi Cheong, Miao, Yun-gen
• Format: Article
• Language: English
• Subjects: Animals; Applied Genetics and Molecular Biotechnology; Bac-to-Bac system; Baculovirus; Biological and medical sciences; Bioreactors; Biotechnology; Bombyx - genetics; Bombyx - metabolism; Bombyx - virology; Bombyx mori; Bombyx mori NPV; Deoxyribonucleic acid; DNA; DNA, Viral - genetics; Dual promoters of Pph and Pp10; E coli; Expression; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes; Genetic engineering; Genetic recombination; Genetic technics; Genetic Vectors - genetics; Hemolymph - virology; Infections; Injection; Larva - genetics; Larva - metabolism; Larva - virology; Larvae; Life Sciences; Manganese; Methods. Procedures. Technologies; Microbial Genetics and Genomics; Microbiology; Modification of gene expression level; Nucleopolyhedrovirus - genetics; Phosphatidylethanolamines; Promoter Regions, Genetic; Reagents; Recombinant baculovirus; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Science; Silk; Silkworm (Bombyx mori L.); Studies; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Transfection; Viral Structural Proteins - genetics; Viruses
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-007-1337-6

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.