大叶补血草Na^+/H^+逆向转运蛋白基因的克隆及序列分析  

Cloning and sequence analysis of a Na^+/H^+ antiporter gene in the halophyte Limonium gmelinii

作　　者：周玲玲[1] 缪建锟[1] 祝建波[1] 曹连莆[2] 

机构地区：[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

出　　处：《草业学报》2009年第5期176-183,共8页Acta Prataculturae Sinica

基　　金：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);石河子大学自然科学与技术创新项目(ZRKX2008034)资助

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 656bp,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。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 LgNHXl were 82.62%, 82.01%, 80.64% and 75.86%identitical in amino acids to the vacuolartyped Na^＋/H^＋ antiporter genes from Atriplex grnelini, Tetragonia tetragonioides, Salicornia europaea and Arabidopsis thaliana, respectively. TMpred prediction showed 11-12 transmembrane domains of LgNHXl. Phylogenetic analysis indicates that LgNHXl 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.

关 键 词：大叶补血草 Na+/H+逆向转运蛋白基因(NHX1) CDNA克隆 RACE 

分 类 号：S54[农业科学—作物学] S688.4