Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand.
Parasitol Res. 2012 Feb;110(2):629-36. doi: 10.1007/s00436-011-2532-z. Epub 2011 Jul 28.
The Cu/Zn superoxide dismutase gene from Wuchereria bancrofti (Cu/Zn WbSOD) was isolated by PCR using degeneracy primers. The complete Cu/Zn WbSOD consisted of 1,032 nucleotides containing 4 exons (477 nucleotides) and 3 introns. The molecular phylogenetic analysis of the Cu/Zn WbSOD gene in comparison with those from other organisms revealed that the gene was classified in the same clade to those of filarial Brugia malayi and Brugia pahangi (bootstrap value at 90). The nucleotide and deduced amino acid sequences of Cu/Zn WbSOD exhibited the similarity to those of intracellular Cu/Zn SOD of B. malayi and B. pahangi. The amino acid comparison of Cu/Zn WbSOD to others revealed that the binding sites and active sites were conserved. The expression of this gene yielded 16.366 kDa in size. After Ni-IDA column purification, the enzyme showed specific activity of 8.5 U/mg and 42.1% yield. The enzyme activity was inhibited when 6 mM KCN was added.
采用简并引物通过 PCR 从班氏吴策线虫（Cu/Zn WbSOD）中分离出 Cu/Zn 超氧化物歧化酶基因。完整的 Cu/Zn WbSOD 由 1032 个核苷酸组成，包含 4 个外显子（477 个核苷酸）和 3 个内含子。与其他生物体的 Cu/Zn WbSOD 基因进行分子系统发育分析表明，该基因与丝虫曼氏吴策线虫和盘尾丝虫（自举值为 90）的基因分类在同一分支。Cu/Zn WbSOD 的核苷酸和推导的氨基酸序列与 B. malayi 和 B. pahangi 的细胞内 Cu/Zn SOD 具有相似性。Cu/Zn WbSOD 与其他氨基酸的比较表明，结合位点和活性位点是保守的。该基因的表达产物大小为 16.366 kDa。经 Ni-IDA 柱纯化后，该酶的比活为 8.5 U/mg，产率为 42.1%。当加入 6 mM KCN 时，酶活性被抑制。
