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细胞色素P450基因CYP9A12的过量表达已被证实与棉铃虫Heliocverpa armigera对拟除虫菊酯的抗性相关。为探明棉铃虫CYP9A12基因的表达调控机理,根据棉铃虫CYP9A12基因cDNA全长的5′-末端核苷酸序列,采用基因组步移方法,获得CYP9 A12的5′-上游区序列(总长为3575bp)。与cDNA序列进行比对,表明在起始密码子上游3bp处有一长为2124bp的内含子。利用NNPP分析软件预测出转录起始位点,与根据CYP9A12全长cDNA序列推测的结果是一致的。TFSEARCH 1.3软件分析转录因子结合位点的结果显示,该序列不仅包含启动子的核心结构序列——TATA-box和CAAT-box,亦包含多个转录因子结合位点,如GATA-1,CdxA,Dfd等。本研究结果为深入研究棉铃虫CYP9A12的表达调控机制及其参与杀虫剂抗性的分子机理奠定了一定基础。
The overexpression of CYP9A12, a cytochrome P450 gene, has been shown to be associated with pyrethroid resistance in Heliocarpa armigera. In order to find out the mechanism of CYP9A12 gene expression in cotton bollworm, 5’-upstream nucleotide sequence of CYP9A12 gene of cotton bollworm was used to obtain 5’-upstream region of CYP9 A12 (3575 bp in length ). Alignment with the cDNA sequence showed that there was a 2124 bp intron at 3bp upstream of the start codon. The NNPP analysis software was used to predict the transcriptional start site, which was consistent with the conjecture based on the full-length CYP9A12 cDNA sequence. TFSEARCH 1.3 software analysis of transcription factor binding sites showed that the sequence contains not only the core structural sequence of the promoter, TATA-box and CAAT-box, but also multiple transcription factor binding sites such as GATA-1, CdxA, Dfd and so on. The results of this study lay a solid foundation for further study on the mechanism of CYP9A12 expression and its molecular mechanism involved in insecticide resistance.