title: A Highly Efficient and Simple Construction Strategy for Producing Recombinant Baculovirus Bombyx mori Nucleopolyhedrovirus.
abstract: The silkworm baculovirus expression system is widely used to produce recombinant proteins. Several strategies for constructing recombinant viruses that contain foreign genes have been reported. Here, we developed a novel defective-rescue BmNPV Bacmid (reBmBac) expression system. A CopyControl origin of replication was introduced into the viral genome to facilitate its genetic manipulation in Escherichia coli and to ensure the preparation of large amounts of high quality reBmBac DNA as well as high quality recombinant baculoviruses. The ORF1629, cathepsin and chitinase genes were partially deleted or rendered defective to improve the efficiency of recombinant baculovirus generation and the expression of foreign genes. The system was validated by the successful expression of luciferase reporter gene and porcine interferon γ. This system can be used to produce batches of recombinant baculoviruses and target proteins rapidly and efficiently in silkworms.
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Xingjian
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Yonglong
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Yinü
authors.initials: Y
authors.lastname: Li
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Haoyang
authors.initials: H
authors.lastname: Li
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yang
authors.affiliation: The Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu Province, China.
authors.forename: Yongzhu
authors.initials: Y
authors.lastname: Yi
authors.affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D012831:Bombyx / Q000821:virology*; D002478:Cells, Cultured / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D007814:Larva / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011679:Pupa / Q000821:virology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D011994:Recombinant Proteins