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草业学报 ›› 2009, Vol. 18 ›› Issue (5): 176-183.

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大叶补血草Na+/H+ 逆向转运蛋白基因的克隆及序列分析

周玲玲1,缪建锟1,祝建波1,曹连莆2*   

  1. 1.石河子大学生命科学学院 农业生物技术重点实验室,新疆 石河子 832000;
    2.石河子大学农学院,新疆 石河子 832000
  • 收稿日期:2008-11-19 出版日期:2009-10-20 发布日期:2009-10-20
  • 作者简介:周玲玲(1966-),女,新疆伊宁人,副教授,在读博士。E-mail:msyzl@163.com
  • 基金资助:
    国家转基因专项“转新型抗旱、耐盐碱等基因的棉花种质资源材料创造”(2008ZX08005-004)和石河子大学自然科学与技术创新项目(ZRKX2008034)资助

Cloning and sequence analysis of aNa+/H+ antiporter gene in the halophyte Limonium gmelinii

ZHOU Ling-ling1, MIAO Jian-kun1, ZHU Jian-bo1, CAO Lian-pu2   

  1. 1.Key Laboratory of Agriculture Biological Technology of College of Life-Science, Shihezi University,
    Shihezi 832000, China;
    2.College of Agriculture, Shihezi University, Shihezi 832000, China
  • Received:2008-11-19 Online:2009-10-20 Published:2009-10-20

摘要: 根据同源序列区域设计简并引物,利用RT-PCR及RACE方法从盐生植物大叶补血草中分离了Na+/H+逆向运输蛋白基因的cDNA(LgNHX1,GenBank登录号EU780457)。LgNHX1的cDNA全长为2 397 bp,5′端非翻译区长度为512 bp, 3′端非翻译区长度为229 bp,其中包括12 bp的poly(A)尾巴,中间的开放读码框为1 656 bp,编码1个550个氨基酸的多肽,推测分子量为61 kD。氨基酸同源性分析表明,LgNHX1与北滨藜、小麦、盐角草和拟南芥液泡Na+/H+逆向运输蛋白的一致性分别为82.62%,82.01%,80.64%和75.86%。预测分析表明,LgNHX1具有11~12个跨膜结构区域。系统发育分析表明该蛋白(LgNHX1)与液泡型的Na+/H+逆向转运蛋白的亲缘关系较近,与质膜型的Na+/H+逆向转运蛋白亲缘关系较远。

Abstract: A vacuolar-typed Na+/H+ antiporter gene was isolated from the halophyte L. gmelinii by RT-PCR and RACE. The LgNHX1 cDNA was 2 397 bp including a 5′untranslated region of 512 bp and a 3′-nontranslated region of 229 bp with poly A, and an open reading frame of 1 656 bp, encoding a predicted polypeptide of 550 amino acids with a predicted molecular mass of 61 kD. It was shown by multiple sequence alignment analysis that the LgNHX1 were 82.62%, 82.01%, 80.64% and 75.86% identitical in amino acids to the vacuolar-typed Na+/H+ antiporter genes from Atriplex gmelini, Tetragonia tetragonioides, Salicornia europaea and Arabidopsis thaliana, respectively. TMpred prediction showed 11-12 transmembrane domains of LgNHX1. Phylogenetic analysis indicates that LgNHX1 is more related to the vacuolar-typed Na+/H+ antiporter, compared with the plasma membrane-type Na+/H+ antiporter, to which it is less related.

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