Cloning,co-expression with an amidating enzyme,and activity of the scorpion toxin BmK ITa1 cDNA in insect cells |
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Authors: | Liu Zhen Yang Guanzhen Li Boliang Chi Chengwu Wu Xiangfu |
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Institution: | (1) State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Shanghiai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, 200031 Shanghai, Peoples Republic of China |
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Abstract: | BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect
cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine α-amidating monooxygenase (PAM) gene.
We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with
coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been
killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression
of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1. |
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Keywords: | BmK ITa1 peptidylglycine α -amidating monooxygenase simultaneous expression C-terminal amidation baculorivus |
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