背角无齿蚌过氧化氢酶基因的克隆及多溴联苯醚对其转录的影响
Cloning of AwCAT from Anodonta woodiana and the Effect of PBDE on Its Transcription
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摘要: 过氧化氢酶（catalase，CAT）是一种广泛存在于各种原核生物和真核生物体内的活性酶，可以将H2O2分解为H2O和O2。为了探讨多溴联苯醚PBDE-47和PBDE-209对背角无齿蚌的胁迫效应，克隆出AwCAT全基因序列，分析PBDE-47和PBDE-209对AwCAT表达的影响。背角无齿蚌AwCAT cDNA全长由1 784个核苷酸组成，开放阅读框包含1 536 bp核苷酸，编码512个氨基酸。AwCAT包含CAT家族高度保守催化位点序列（61FDRERIPERVVHAKGAGA77）和血红素配体信号序列（351RLFSYSDTH358）2个特征性标签序列。与对照组相比，在6.25、12.5、25、50和100 μg·L-1的PBDE-47处理组中，AwCAT mRNA水平分别增加了66.66%（P<0.05）、1.35倍（P<0.05）、1.54倍（P<0.05）、1.97倍（P<0.01）和2.39倍（P<0.01）以上；在10、20、40、80和160 μg·L-1的PBDE-209处理组中，AwCAT mRNA水平分别增加了7.84%、35.38%、61.53%（P<0.05）、1.03倍（P<0.05）和1.09倍（P<0.05）以上。与对照组相比，PBDE-47处理后鳃中AwCAT mRNA水平显著升高；PBDE-209处理后鳃中AwCAT mRNA水平增加了85.71%（P<0.05）以上。以上结果表明，PBDE-47和PBDE-209处理对背角无齿蚌AwCAT表达具有明显的诱导作用，有助于提高机体对H2O2的清除能力和抗氧化能力。Abstract: Catalase (CAT) is a major reactive oxygen species (ROS) scavenger enzyme that plays a significant role in the antioxidant defense mechanism of organisms by reducing toxic hydrogen peroxide molecules into a nontoxic form of oxygen and water with a high turnover rate. Polybrominated diphenyl ethers (PBDEs) are currently important components in flame retardants. Excess PBDEs deposited in organism could catalyze the production of ROS. In current study, one complete CAT sequence was isolated from Anodonta woodiana and named AwCAT . The full-length AwCAT cDNA of A. woodiana consists of 1 784 nucleotides. The open reading frame contains 1 536 bp nucleotides and encodes 512 amino acids. Compared with that of control group, AwCAT mRNA level of hepatopancreas increased more than 66.66% (P<0.05), 1.35 times (P<0.05), 1.54 times (P<0.05), 1.97 times (P<0.01) and 2.39 times (P<0.01) in the 6.25, 12.5, 25, 50 and 100 μg·L-1 of PBDE-47 treated groups, respectively; AwCAT mRNA level of hepatopancreas increased more than 7.84%, 35.38%, 61.53% (P<0.05), 1.03 times (P<0.05) and 1.09 times (P<0.05) in the 10, 20, 40, 80 and 160 μg·L-1 of PBDE-209 treated group, respectively. Compared with that of control group, AwCAT mRNA level of gill significantly increased in the PBDE-47 treated groups. Compared with that of control group, AwCAT mRNA level of gill increased more than 85.71% (P<0.05) in the PBDE-209 treated goups. These results indicate that up-regulations of AwCAT expression of hepatopancreas and gill in the freshwater bivalve A. woodiana are contributed to eliminate stress derived from PBDE-47 and PBDE-209 challenge.
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Key words:
- polybrominated diphenyl ethers /
- Anodonta woodiana /
- antioxidative ability /
- PBDE-47 /
- PBDE-209 /
- AwCAT
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