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日本囊对虾中CYP4两种同工型的鉴定及其对苯并[a]芘的mRNA表达谱响应

Identification of two isoforms of CYP4 in Marsupenaeus japonicus and their mRNA expression profile response to benzo[a]pyrene.

作者信息

Zheng Jinbin, Mao Yong, Qiao Yin, Shi Zhuangzhuang, Su Yongquan, Wang Jun

机构信息

College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China.

College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, China.

出版信息

Mar Environ Res. 2015 Dec;112(Pt A):96-103. doi: 10.1016/j.marenvres.2015.09.012. Epub 2015 Oct 9.

Abstract

CYP4 enzymes are essential components of cellular detoxification systems and play important roles in monitoring persistent organic pollutants in marine environments. However, there are few studies on CYP4 in shrimp. In this study, two CYP4 isoforms, CYP4V28 and CYP4V29, were cloned from Marsupenaeus japonicus for the first time, and the tissue distributions and mRNA expression profile in response to benzo[a]pyrene (B[a]P) were analyzed by quantitative real-time PCR (QRT-PCR). The full lengths of CYP4V28 and CYP4V29 were 1771 bp and 1647 bp respectively, with deduced amino acid sequences of 511 and 515 amino acids. The two CYP4s were predominantly expressed in the hepatopancreas and weakly expressed in other six tested tissues. As demonstrated by QRT-PCR, the mRNA levels of the two CYP4s show both a time- and dose-dependent response to B[a]P. The mRNA expression levels of CYP4V28 and CYP4V29 peaked at 12 h and 6 h respectively, and the peak level exhibited a tendency of positive correlation with the concentration of B[a]P. This study provides clues for further elucidating the function and regulation mechanisms of the two CYP4s in M. japonicas and evaluating of the biomarker potential of the two CYP4 isoforms.

摘要

细胞色素P450 4(CYP4)酶是细胞解毒系统的重要组成部分,在监测海洋环境中的持久性有机污染物方面发挥着重要作用。然而,关于虾类中CYP4的研究较少。在本研究中,首次从日本囊对虾中克隆出两种CYP4亚型,即CYP4V28和CYP4V29,并通过定量实时荧光定量PCR(QRT-PCR)分析了它们在不同组织中的分布以及对苯并[a]芘(B[a]P)的mRNA表达谱。CYP4V28和CYP4V29的全长分别为1771 bp和1647 bp,推导的氨基酸序列分别为511和515个氨基酸。这两种CYP4主要在肝胰腺中表达,在其他六个检测组织中表达较弱。QRT-PCR结果表明,这两种CYP4的mRNA水平对B[a]P呈现出时间和剂量依赖性反应。CYP4V28和CYP4V29的mRNA表达水平分别在12小时和6小时达到峰值,且峰值水平与B[a]P浓度呈正相关趋势。本研究为进一步阐明这两种CYP4在日本囊对虾中的功能和调控机制以及评估这两种CYP4亚型作为生物标志物的潜力提供了线索。

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