FEMS Microbiology Letters (2010) 302, 46-51
Furong Tan, Aiping Zheng, Jun Zhu, Lingxia Wang, Shuangcheng Li, Qiming Deng, Shiquan Wang, Ping Li and Xueming Tang (2010)
Rapid cloning, identification, and application of one novel crystal protein gene cry30Fa1 from Bacillus thuringiensis
FEMS Microbiology Letters 302 (1), 46-51
Abstract: In this study, a fast and efficient strategy has been developed for identifying and isolating novel cry genes from Bacillus thuringiensis by combining the PCR-restriction fragment length polymorphism and the single-oligonucleotide nested-PCR method. Using this method, one novel holotype cry gene, cry30Fa1, encoding a polypeptide of 687 amino acid residues with a molecular mass of 77.1 kDa, 74% identical to Cry30Aa1, was cloned from the B. thuringiensis strain BtMC28. Furthermore, the cry30Fa1 gene was successfully expressed in Escherichia coli BL21 (DE3). The Cry30Fa1 proteins, isolated from the cultures of recombinant E. coli, had remarkable insecticidal effects against Plutella xylostella and Aedes aegypti with LC50 at 6.477 and 15.359 μg mL-1, respectively. Our results strongly suggest that this strategy is highly efficient and advantageous in terms of rapid cloning of holotype cry genes that have minimal identity to known genes. The cloning of the cry30Fa1 gene would be useful in the resources of the insecticidal crystal genes and may serve as an alternative choice of an insecticide for potential problems associated with insect resistance.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Aiping Zheng, Ping Li
Research topic(s) for pests/diseases/weeds:
biocontrol - natural enemies
Research topic(s) for beneficials or antagonists:
general biology - morphology - evolution
molecular biology - genes
Pest and/or beneficial records:
| Beneficial | Pest/Disease/Weed | Crop/Product | Country | Quarant.
|
|---|---|---|---|---|
| Plutella xylostella | ||||
| Aedes aegypti | ||||
| Bacillus thuringiensis Cry30-toxin (entomopathogen) | Plutella xylostella | |||
| Bacillus thuringiensis Cry30-toxin (entomopathogen) | Aedes aegypti |
