背角无齿蚌AwPrx4基因克隆与PFOS和PFOA胁迫表达分析

Effects of PFOS and PFOA on the AwPrx4 Expression in Freshwater Bivalve Anodonta woodiana

为了探讨全氟辛烷磺酸(PFOS)和全氟辛酸(PFOA)对背角无齿蚌的胁迫效应,将背角无齿蚌随机分为对照组、PFOS处理组和PFOA处理组;同时克隆出Aw Prx4全基因序列,分析PFOS和PFOA对Aw Prx4表达的影响。Aw Prx4全长cDNA开放阅读框由588 bp核酸组成,编码196个氨基酸,包含FYPLDFTFACPTEI和GEVCPA两个标签序列。肝胰腺Aw Prx4 mRNA水平在6.25 mg/L、12.5 mg/L和25 mg/L的PFOS处理组中的上调效应呈时间和剂量依赖模式;与对照组相比,在50 mg/L和100 mg/L PFOS处理中Aw Prx4 m RNA水平分别增加了6.11倍(p<0.01)和6.68倍(p<0.01)。与对照组相比,PFOA处理组中(50 mg/L,100 mg/L和200 mg/L)肝胰腺Aw Prx4 mRNA水平增加了37.05%以上;在PFOA处理组中(400 mg/L和800 mg/L),肝胰腺Aw Prx4表达水平在整个实验过程中分别增加了1.36倍(p<0.05)和75.76%(p<0.05)。与对照组相比,PFOS和PFOA处理后腮中Aw Prx4表达水平在整个实验过程中分别增加了64.64%(p<0.05)和69.69%。与对照组相比,PFOS和PFOA处理后血淋巴中Aw Prx4表达水平分别增加了37.11%和38.24%以上。这些研究结果为认识其他物种Prx4基因的结构和功能提供参考,同时为揭示双壳类动物对抗持久性有机污染物的胁迫效应提供了理论依据。

To explore the stress effects of perfluorooctane sulfonic acid(PFOS)and perfluorooctanoic acid(PFOA)on toothless mussels,the Anodonta woodiana were randomly divided into control group,PFOS treatment group and PFOA treatment group;at the same time,the complete gene sequence of Aw Prx4 was cloned and the effects of PFOS and PFOA on the expression of Aw Prx4 were analyzed.The full-length cDNA of Aw Prx4 had an open reading frame(ORF)of 588 bp encoding 196 amino acids.Two highly conserved Prxs signature motifs were observed in deduced amino acid sequence,one was FYPLDFTFACPTEI,and the other was GEVCPA.Compared with that of control group,up-regulation of Aw Prx4 mRNA level showed time and dose-dependent pattern in the 6.25 mg/L,12.5 mg/L,25 mg/L of PFOS treated groups.Aw Prx4 m RNA level increased more than 6.11 times(p<0.01)and6.68 times(p<0.01)in the 50 mg/L,100 mg/L of PFOS treated group incontrasted with that ofcontrol group,respectively.Compared with the control group,the hepatopancreal AwPrx4 mRNA levels in the PFOA treated group(50 mg/L,100 mg/L and 200 mg/L)increased by more than 37.05%.Compared with that of control group,Aw Prx4 m RNA level increased 1.36 times(p<0.05)and 75.76%(p<0.05)in the 400 mg/L and 800 mg/L of PFOA treated groups.Compared with that of control group,Aw Prx4 mRNA level of gill increased 64.64%(p<0.05)and 69.69%in the PFOS treated groups and PFOA treated groups.Compared with that of control group,Aw Prx4 m RNA level of hemocytes increased 37.11%(p<0.05)and 38.24%in the PFOS treated groups and PFOA treated groups.These results provide references for understanding the structure and function of the PRx4 gene in other species,and provide theoretical basis for revealing the stress effects of bivalves against persistent organic pollutants.