福寿螺细胞色素P450基因CYP3192A1的克隆与表达分析

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...

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Bibliographic Details
Published in南方水产科学 Vol. 13; no. 1; pp. 66 - 75
Main Author 张文领 牟希东 胡隐昌 宋红梅
Format Journal Article
LanguageChinese
Published 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,广东 广州510380 2017
上海海洋大学水产与生命学院,上海201306%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,广东 广州510380
Subjects
四聚乙醛
福寿螺
细胞色素P450
表达分析
细胞色素P450
expression analysis
四聚乙醛
Cytochrome P450
表达分析
福寿螺
metaldehyde
Pomacea canaliculata
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ISSN2095-0780
DOI10.3969/j.issn.2095-0780.2017.01.009

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Summary: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（P〈0.05）,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。
Bibliography:The study investigates the structure and expression features of Pomacea canaliculata cytochrome P450,and explores the correlation of metaldehyde treatment and cytochrome P450 expression level. A full-length c DNA of CYP3192A1 from P. canaliculata with total length of 2 523 bp was obtained by RACE,including an open reading frame（ ORF） encoding 517 amino acids,which had a heme-binding domain F＊＊G＊＊＊ C＊ G,helix K（ E＊＊R）,helix I（ AG＊ ET）,helix C（ W＊＊＊R） and trans-membrane helix（ 12～ 34 amino acids）. Blast and phylogenetic analyses suggest P. canaliculata CYP3192A1 is a member of a novel CYP450 family,with high similarity with CYP3 A subfamily. The RT-q PCR suggests that CYP3192A1 has gender-specific and tissue specific expression（ P 〈0. 05）. Expression in female P. canaliculata was higher than in males; expression amounts in intestinal and gill were the highest,followed by those in liver and stomach,and were the lowest in heart and pleopod. The influence of metaldehyde to CYP3192A1 expression shows a trend of ind
ISSN:2095-0780
DOI:10.3969/j.issn.2095-0780.2017.01.009