引用本文:	张楠楠,潘卫东,崔玉琳,秦松,薛乐勋.杜氏盐藻磷酸烯醇式丙酮酸羧化酶基因的克隆和分析[J].海洋科学,2011,35(12):42-47.

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杜氏盐藻磷酸烯醇式丙酮酸羧化酶基因的克隆和分析
张楠楠¹, 潘卫东², 崔玉琳³, 秦松⁴, 薛乐勋¹
1.郑州大学生物工程系细胞生物学研究室;2.郑州大学基础医学院微生物学与免疫学教研室;3.中国科学院海洋研究所;4.中国科学院烟台海岸带研究所

摘要:

为研究杜氏盐藻(Dunaliella salina)磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因的功能, 根据莱茵衣藻(Chlamydomonas reinhardtii)、拟南芥(Arabidopsis thaliana)、花生(Arachis hypogaea)等真核生物PEPC 基因高度保守序列, 设计一对简并引物, 通过RT-PCR 的方法获得杜氏盐藻PEPC 基因部分序列, 然后采用RACE 的方法分别克隆到5′端和3′端序列, 拼接后得到全长cDNA, 其长度为3 523bp, 包含2 949 bp 的完整开放阅读框, 编码982 个氨基酸, 相对分子质量为110560.5。氨基酸序列与已知物种PEPC 序列的同源性依次为: Chlamydomonas reinhardtii 69%, Chlorellavariabilis 55%, Ostreococcus tauri 50%和Ostreococcus lucimarinus CCE9901 49%, 表明所克隆的序列确为杜氏盐藻PEPC cDNA 序列。

关键词: 杜氏盐藻(Dunaliella salina) 磷酸烯醇式丙酮酸羧化酶基因克隆序列分析

DOI：

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Cloning and analysis of phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina

Abstract:

To investigate the function of phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina, a pair of degenerate primers was designed according to the conserved motifs of the PEPC of Chlamydomonas reinhardtii, Arabidopsis thaliana, and Arachis hypogaea. A cDNA fragment was obtained from green alga Dunaliella salina through RT-PCR, and then the full length of the cDNA was isolated by 3’ and 5’RACE. The isolated cDNA sequence was 3523 bp in length with a 2949 bp coding region that encoded 982 amino acid residues with the predicted relative molecular mass of 110560.5 dolton. In addition, homology analysis showed that PEPC of D. salina was highly similar to that of C. reinhardtii(69%), Chlorella variabilis(55%), Ostreococcus tauri(50%), and O. lucimarinus CCE9901(49%), suggesting that the cDNA isolated from Dunaliella salina was PEPC-encoding.

Key words: Dunaliella salina PEPC gene cloning sequence analysis