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采用同源克隆和RACE技术,从孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant中克隆到细胞色素P450CYP9Z401基因的c DNA全序列(Gen Bank number:KP164813)。c DNA全长1722 bp,包含3’非编码区域(UTR)为86 bp和5’UTR为49 bp,开放阅读框(ORF)长1587 bp,编码528个氨基酸。预测的分子量为61.27 k D,理论等电点为6.58,无信号肽结构,在2-20氨基酸处存在跨膜螺旋。该基因拥有细胞色素P450特征序列螺旋C、螺旋K和Meander区域,另外还有血红素结合区(1个氨基酸差异)和螺旋I区(2个氨基酸差异)。与其他昆虫的CYP9家族基因比较,与赤拟谷盗Tribolium castaneum同源性最高,为47%,系统发育分析也表明两者的亲缘关系最近。CYP9Z401基因的克隆和比较分析为进一步深入研究孟氏隐唇瓢虫的细胞色素P450基因功能及其进化具有重要意义。
The c DNA sequence of cytochrome P450CYP9Z401 (GenBank number: KP164813) was cloned from Cryptolaemus montrouzieri Mulsant using homologous cloning and RACE techniques. The length of c DNA was 1722 bp, including 86 bp in 3 ’untranslated region (UTR) and 49 bp in 5’UTR. The open reading frame (ORF) was 1587 bp in length and encoded 528 amino acids. The predicted molecular weight was 61.27 kD and the theoretical isoelectric point was 6.58. There was no signal peptide structure, and there was a transmembrane helix at 2-20 amino acids. The gene has the cytochrome P450 signature sequence helix C, helix K and the Meander region, in addition to the heme binding region (1 amino acid difference) and the helix I region (2 amino acid differences). Compared with CYP9 family genes of other insects, it has the highest homology with Tribolium castaneum of 47%. Phylogenetic analysis also shows that the two have the closest genetic relationship. The cloning and comparative analysis of CYP9Z401 gene is of great significance for further study on the function and evolution of cytochrome P450 gene in ladybugs.