Comparative studies of lepidopteran baculovirus-specific protein FP25K: development of a novel Bombyx mori nucleopolyhedrovirus-based vector with a modified fp25K gene.
Lepidopteran baculovirus-specific protein FP25K performs many roles during the infection cycle, including functions in the production of occlusion bodies (OBs) and budded viruses (BVs), oral infection, and postmortem host degradation. To explore the common and specific functions of FP25K proteins among lepidopteran baculoviruses, we performed comparative analyses of FP25K proteins from group I and group II nucleopolyhedroviruses (NPVs) and granulovirus (GV). Using recombinant Bombyx mori NPVs (BmNPVs), we showed that the FP25Ks from NPVs were able to eliminate all the phenotypic defects observed in an infection with a BmNPV mutant lacking functional fp25K but that FP25K from GV did not show abilities to recover oral infectivity and postmortem host degradation. We also observed that introduction of Autographa californica multiple NPV (AcMNPV) fp25K into the BmNPV genome enhanced OB and BV production. According to these results, we generated a novel BmNPV-based expression vector with AcMNPV fp25K and examined its potential in BmN cells and B. mori larvae. Our results showed that the introduction of AcMNPV fp25K significantly increases the expression of foreign gene products in cultured cells and shortens the time for obtaining the secreted recombinant proteins from larval hemolymph.
Nakanishi
Tadashi
T
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Goto
Chie
C
Kobayashi
Michihiro
M
Kang
Wonkyung
W
Suzuki
Takehiro
T
Dohmae
Naoshi
N
Matsumoto
Shogo
S
Shimada
Toru
T
Katsuma
Susumu
S
D000818:Animals; D001709:Biotechnology / Q000379:methods; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D025722:Granulovirus / Q000235:genetics / Q000502:physiology*; D007915:Lepidoptera / Q000821:virology*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000502:physiology*; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis
D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins