Open AccessRapid detection of vip1 ‐ type genes from B acillus cereus and characterization of a novel vip binary toxin gene
Author(s) -
Yu Xiumei,
Liu Tao,
Liang Xiaoxing,
Tang Changqing,
Zhu Jun,
Wang Shiquan,
Li Shuangcheng,
Deng Qiming,
Wang Linxia,
Zheng Aiping,
Li Ping
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02409.x
Subject(s) - biology , pore forming toxin , gene , bacillus cereus , spodoptera , microbiology and biotechnology , restriction fragment length polymorphism , helicoverpa armigera , escherichia coli , cereus , toxin , genetics , polymerase chain reaction , botany , recombinant dna , lepidoptera genitalia , bacteria , microbial toxins
A PCR –restriction fragment length polymorphism ( PCR – RFLP ) method for identifying vegetative insecticidal protein ( vip ) 1 ‐ type genes from B acillus cereus was developed by designing specific primers based on the conserved regions of the genes to amplify vip1 ‐ type gene fragments. PCR products were digested with endonuclease AciI , and four known vip1 ‐type genes were identified. Vip1Ac and vip1Aa ‐type genes appeared in 17 of 26 B . cereus strains. A novel vip1 ‐type gene, vip1Ac1 , was identified from B . cereus strain HL 12. The vip1Ac1 and vip2Ae3 genes were co‐expressed in E scherichia coli strain B L21 by vector pCOLAD uet‐1. The binary toxin showed activity only against A phis gossypii ( H omoptera), but not for C oleptera ( T enebrio molitor , H olotrichia oblita ), L epidoptera ( S podoptera exigua , H elicoverpa armigera , and C hilo suppressalis ), Diptera ( C ulex quinquefasciatus ). The LC 50 of this binary toxin for A . gossypii is 87.5 (34.2–145.3) ng mL −1 . This is probably only the second report that Vip 1 and Vip 2 binary toxin shows toxicity against homopteran pests. The PCR – RFLP method developed could be very useful for identifying novel Vip 1– Vip 2‐type binary toxins, and the novel binary toxins, Vip1Ac1 and Vip2Ae3 , identified in this study may have applications in biological control of insects, thus avoiding potential problems of resistance.