九孔鲍铜锌超氧化物歧化酶的克隆、表征及三丁基锡暴露响应
Cloning, characterization and TBT exposure response of CuZn superoxide dismutase from Haliotis diversicolor supertexta.
作者信息
Zhang Kefeng, Wang Guodong, Zou Zhihua, Jia Xiwei, Wang Shuhong, Lin Peng, Chen Yun, Zhang Ziping, Wang Yilei
机构信息
The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, Fujian, China.
出版信息
Mol Biol Rep. 2009 Mar;36(3):583-94. doi: 10.1007/s11033-008-9217-4. Epub 2008 Mar 24.
The full-length cDNA and genomic DNA of a cytoplasmic copper, zinc superoxide dismutase (CuZn-sod) were cloned from the hepatopancreas of small abalone Haliotis diversicolor supertexta by RT-PCR, RACE and TAIL PCR. The full-length cytoplasmic CuZn-sod cDNA (designated sasod) comprises 984 bp. Its ORF encodes a polypeptide of 154 amino acids with a predicted molecular mass of 15.7 kDa and theoretical isoelectric point of 6.30. The deduced amino acid (designated saSOD) shares a common consensus pattern with the SODs of vertebrate and invertebrate animals. The full-length sasod genomic DNA comprises 5,574 bp, containing five exons and four introns. The splice donor and acceptor sequence of the four introns is 5'GT-AG3'. Real time quantitative PCR analysis revealed that sasod expression level in hepatopancreas of small abalone was no significant difference at 2, 6, 48 and 192 h post TBT exposure (P > 0.05). However, the sasod expression level at 12 and 24 h post TBT exposure was decreased significantly (P < 0.05).
通过逆转录聚合酶链反应（RT-PCR）、cDNA末端快速扩增技术（RACE）以及热不对称交错PCR（TAIL PCR），从小皱纹盘鲍（Haliotis diversicolor supertexta）的肝胰腺中克隆出细胞质铜锌超氧化物歧化酶（CuZn-sod）的全长cDNA和基因组DNA。细胞质CuZn-sod的全长cDNA（命名为sasod）包含984个碱基对。其开放阅读框编码一个由154个氨基酸组成的多肽，预测分子量为15.7 kDa，理论等电点为6.30。推导的氨基酸序列（命名为saSOD）与脊椎动物和无脊椎动物的超氧化物歧化酶具有共同的共有模式。sasod基因组DNA全长5574个碱基对，包含5个外显子和4个内含子。4个内含子的剪接供体和受体序列为5'GT-AG3'。实时定量PCR分析显示，在三丁基锡（TBT）暴露后2、6、48和192小时，小皱纹盘鲍肝胰腺中sasod的表达水平无显著差异（P>0.05）。然而，在TBT暴露后12和24小时，sasod的表达水平显著降低（P<0.05）。
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