| 摘要: |
| 利用已报道的FeSOD保守区域, 设计简并引物, 通过简并PCR 的方法获得了雨生红球藻(Haematococcus pluvialis)FeSOD(HpFeSOD)cDNA 的部分序列, 然后采用RACE 方法分别克隆到5′端和3′端。拼接后得到HpFeSODcDNA 全长, 该基因全长为1 138 bp, ORF 为684 bp, 编码227 个氨基酸。把已经得到的HpFeSOD序列推导成氨基酸序列与一些已知物种的FeSOD氨基酸序列相比较, 雨生红球藻FeSOD与杜氏藻、衣藻、团藻、石莼、颤藻的同源性为89%、83%、78%、70%和63%。分子系统学分析表明, 雨生红球藻FeSOD与杜氏藻FeSOD聚在一起, 介于真菌与高等植物之间并且真核微藻FeSOD与高等植物的同源性更高, 推测其在进化上与高等植物的亲源关系更近。 |
| 关键词: 雨生红球藻(Haematococcus pluvialis) 超氧化物歧化酶 基因克隆 序列分析 |
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| 基金项目:山东省杰出青年学者基金项目(JQ200914); 国家自然科学基金项目(31000037); 中国科学院知识创新工程计划项目(KSCX2-YW-G-073, KZCX2-YW-216, KZCX2-YW-209) |
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| Cloning and sequence analysis of a iron superoxide dismutase from Haematococcus pluvialis |
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| Abstract: |
| A pair of degenerate primers was designed to amplify specific DNA fragment of FeSOD using cDNA of Haematococcus pluvialis from the homologous sequences. The middle interesting cDNA fragment was obtained by RT-PCR. The full length of HpFeSOD cDNA was obtained by 5′RACE and 3′RACE. The clone contains 1138 bp nucleotides with an open reading frame (ORF) of 684 bp comprising 227 amino acid residues. The sequence shares a high homologue with the following FeSOD: Dunaliella salina, 89%, Chlamydomonas reinhardtii, 83%, Volvox carteri f. nagariensis, 78%, Ulva fasciata, 70%, and Cyanobium sp. PCC 7001, 63%. Phylogenetic analysis shows that the FeSOD from H. pluvialis and D. salina are clustered together with genes between fungi and higherplants. The FeSOD of eukaryotic microalgae and higher plants may arise through gene duplication events independently from acommon ancestor. |
| Key words: Haematococcus pluvialis FeSOD gene cloning sequence analysis |
