Premium
Identification of vip3A ‐type genes from Bacillus thuringiensis strains and characterization of a novel vip3A ‐type gene
Author(s) -
Liu J.,
Song F.,
Zhang J.,
Liu R.,
He K.,
Tan J.,
Huang D.
Publication year - 2007
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02217.x
Subject(s) - bacillus thuringiensis , biology , gene , spodoptera , restriction fragment length polymorphism , genetics , helicoverpa armigera , homology (biology) , polymerase chain reaction , microbiology and biotechnology , lepidoptera genitalia , botany , bacteria , recombinant dna
Aims: To search for novel Vip3A proteins for controlling insect pests. Methods and Results: A pair of universal primers was designed based on the conserved regions of five vip3A genes. Amplified products were digested with the Hin dIII and Eco R enzymes so as to confirm different restriction fragment length polymorphism (RFLP) patterns used to identify vip3A ‐type genes. The vip3A gene types of 606 Bacillus thuringiensis strains were screened and three patterns of RFLP were successfully identified. Two novel vip3A genes were found and one of these, vip3Aa19 , was further characterized and its product was confirmed toxic to Spodoptera exigua , Helicoverpa armigera and Plutella xylostella larvae. Partial sequences of another novel vip3A‐ type gene were obtained that shared 83% homology with that of the vip3Af1 gene. Conclusions: A polymerase chain reaction (PCR)‐RFLP system we developed could be used for identifying novel vip3A‐ genes from B. thuringiensis strains. A novel Vip3A protein was found to have a broader insecticidal spectrum. Significance and Impact of the Study: The reported method is a powerful tool to find novel Vip3A proteins from large‐scale B. thuringiensis strains. The novel Vip3A protein may be used to control insect pests or resistant insect pests by constructing genetically engineered strains or transgenic plants.