Coexpression of Cry48Aa/Cry49Aa Toxin from Lysinibacillus sphaericus in Crystal-minus B. thuringiensis and Activity against Mosquito Larvae

doi:10.16409/j.cnki.2095-039x.2015.06.019

Abstract

Abstract: The Cry48Aa/Cry49Aa, two-component toxin, shows high toxicity against Culex species, including binary toxin BinA/BinB-resistant Culex larvae. Being a potential novel mosquitocidal toxin, however, the low expression of Cry48Aa in the wild strain limits its development and application. In this work, the promoter of cry48Aa was replaced and cry48Aa and cry49Aa were coexpressed in Bacillus thuringiensis. The expression of recombinant proteins were detected by SDS-PAGE, visualized as a 120 (Cry48Aa) and a 53 kDa (Cry49Aa) bands. The expression amount of cry48Aa was low in its own promoter, which however, was significantly increased after the replacement of its promoter with the strong promoter Pcry1Aa but was still lower than the expression of Cry49Aa. Both the recombinant strains BMB171-pBU-cry48Aa-cry49Aa and BMB171-pBU-Pcry1Aa-cry48Aa- cry49Aa produced bipyramidal and squared crystals. The bioassay result showed that the two strains were effective against the 4th instar larvae of Bin-susceptible and -resistant C. quinquefasciatus, with the LC₅₀ of 3.05 and 3.45, and 0.65 and 0.82 μL/mL, respectively.

Key words: Lysinibacillus sphaericus, Cry48Aa/Cry49Aa, expression, Culex quinquefasciatus, mosquitocidal activity

CLC Number:

S476.11

GUO Qingyun, CAI Quanxin, HU Xiaomin, YAN Jianping, YUAN Zhiming. Coexpression of Cry48Aa/Cry49Aa Toxin from Lysinibacillus sphaericus in Crystal-minus B. thuringiensis and Activity against Mosquito Larvae[J]. journal1, 2015, 31(6): 950-955.

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