福寿螺细胞色素P450基因CYP3192A1的克隆与表达分析

发布时间：2018-06-07 01:15

本文选题：福寿螺 + 细胞色素P　；参考：《南方水产科学》2017年01期

【摘要】：文章分析了福寿螺(Pomacea canaliculata)细胞色素P450(CYPs)的结构、表达特征及四聚乙醛代谢与细胞色素P450表达水平的相关性。应用c DNA末端快速扩增技术(rapid amplification of c DNA ends,RACE)克隆获得福寿螺CYPs基因c DNA全长2 523 bp,命名为CYP3192A1,含1个开放阅读框(ORF),编码517个氨基酸,具有血红素结合区F**G***C*G、K螺旋(E**R)、I螺旋(AG*ET)、C螺旋(W***R)等高度保守序列及跨膜螺旋(12~34号氨基酸)。聚类分析显示福寿螺CYP3192A1是细胞色素P450新家族成员,与CYP3A亚家族序列同源性较高。荧光定量PCR(real time quantitative PCR,RT-q PCR)分析显示,福寿螺CYP3192A1基因表达量存在显著的性别及组织差异性(P0.05),表现为雌性表达量高于雄性,肠和鳃表达量最丰富,肝和胃次之,心和腹足中少量表达;四聚乙醛对福寿螺CYP3192A1表达的影响为"低浓度诱导,高浓度诱导期缩短并提前"。结果表明四聚乙醛处理能够显著诱导福寿螺CYP3192A1表达,与CYP3192A1表达量增加紧密相关,福寿螺CYP3192A1过表达可能在四聚乙醛代谢抗性中起重要作用。
[Abstract]:The structure and expression characteristics of cytochrome P450 CYPsof Pomacea canaliculata and the correlation between the metabolism of tetrapolyacetaldehyde and the expression level of cytochrome P450 were analyzed. The c DNA of CYPs gene was cloned by rapid amplification of c DNA endsrase (RACEE) and named CYP31919A1, which contains an open reading frame and encodes 517 amino acids. It has highly conserved sequences such as heme binding region (FG) and transmembrane helix (12 ~ 34 amino acid). Cluster analysis showed that CYP3192A1 was a new member of cytochrome P450 family and had high homology with CYP3A subfamily. Fluorescence quantitative PCR(real time quantitative PCR RT-Q PCR analysis showed that there were significant differences in the expression of CYP3192A1 gene between male and female (P 0.05). The expression of CYP3192A1 in female was higher than that in male, the most abundant in intestine and Gill, the second in liver and stomach, and a small amount in heart and foot. The effect of tetrapolyacetaldehyde on the expression of CYP3192A1 in snail was "induced by low concentration, shortened and advanced by high concentration". The results showed that tetramer acetaldehyde treatment could significantly induce the expression of CYP3192A1, which was closely related to the increase of CYP3192A1 expression. The overexpression of CYP3192A1 might play an important role in the metabolic resistance of tetraoxal.
【作者单位】：中国水产科学研究院珠江水产研究所农业部热带亚热带水产资源利用与养殖重点实验室;上海海洋大学水产与生命学院;
【基金】：国家自然科学基金项目(31300468) 农业部农业入侵防治项目(2130108)
【分类号】：S917.4

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