caa a22 4500 605504768 CHVBK 20210128100618.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6846-0 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6846-0 Improved insecticidal activity of a recombinant baculovirus expressing spider venom cyto-insectotoxin [Elektronische Daten] [M. Ali, Tatsuya Kato, Enoch Park] Baculoviruses have a long history of safe use as specific, environmentally friendly insecticides that provide alternatives to chemical pesticides for controlling insect pests. However, their use has been limited by several factors, particularly their slow pathogenicity. In this study, we constructed a recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) and an Autographa californica multiple nucleopolyhedrovirus (AcMNPV) that expressed an insect-specific cyto-insectotoxin (Cit1a) from the venom of the central Asian spider Lachesana tarabaevi. Cit1a is a comparatively long linear cytolytic molecule that contains a predicted α-helix structure composed of two short membrane-acting antimicrobial peptides (MAMPs) that are joined together in a "head-to-tail” shape. Cit1a fused to polyhedrin gene (polh) (polh-cit1a) was expressed in the nuclei as polyhedra in silkworm larvae, Bm5 and Sf9 cells. An early death of Bm5 and Sf9 cells by recombinant BmNPV/Polh-Cit1a and AcMNPV/Polh-Cit1a was observed compared with control viruses that lacked the toxin gene. The infected cells showed a loss of cytoplasm, membrane integrity, and structural changes, suggesting that recombinant baculovirus-infected cells were killed by the necrosis caused by Cit1a. In addition, the BmNPV/Polh-Cit1a showed a significant reduction in the median lethal time (LT50) against silkworm larvae compared with those of control BmNPV that lacked the cit1a gene. Springer-Verlag Berlin Heidelberg, 2015 Cyto-insectotoxin nationallicence Recombinant baculovirus nationallicence Pathogenicity nationallicence Cytotoxicity nationallicence Bombyx mori nationallicence Ali M. Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut Kato Tatsuya Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut Park Enoch Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10261-10269 0175-7598 99:23<10261 2015 99 253 https://doi.org/10.1007/s00253-015-6846-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6846-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ali M. Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut NATIONALLICENCE 700 1- Kato Tatsuya Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut NATIONALLICENCE 700 1- Park Enoch Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Suruga-ku, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/23(2015-12-01), 10261-10269 0175-7598 99:23<10261 2015 99 253