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A Novel Negative Regulatory Factor for Nematicidal Cry Protein Gene Expression in Bacillus thuringiensis  

Yu, Ziquan (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Bai, Peisheng (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Ye, Weixing (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Zhang, Fengjuan (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Ruan, Lifang (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Yu, Ziniu (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Sun, Ming (State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University)
Publication Information
Journal of Microbiology and Biotechnology / v.18, no.6, 2008 , pp. 1033-1039 More about this Journal
Abstract
A 3-kb HindIII fragment bearing the cry6Aa2 gene and the adjacent and intergenic regions was cloned from Bacillus thuringiensis strain YBT-1518. Two open reading frames (ORFs), namely, orf1 (termed cry6Aa2) and orf2 that were separated by an inverted-repeat sequence were identified. orf1 encoded a 54-kDa protein that exhibited high toxicity to the plant-parasitic nematode Meloidogyne hapla. The orf2 expression product was not detected by SDS-PAGE, but its mRNA was detected by RT-PCR. The orf2 coexpressed with orf1 at a high level in the absence of the inverted-repeat sequence, whereas, the expression level of otfl was decreased. When orf2 was mutated, the level of orf1 expression was enhanced obviously. In conclusion, the inverted-repeat sequence disturbs orf2 expression, and the orf2 downregulates orf1 expression. This is an example of novel negative regulation in B. thuringiensis and a potential method for enhancing the expression level of cry genes.
Keywords
Bacillus thuringiensis; nematicidal cry gene; expression; negative regulation;
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