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采用简并引物和RACE等技术从荷花中分离出液泡膜型Na+/H+逆向转运蛋白基因NHX1的全长cDNA序列,命名为NnNHX1。该基因全长2237bp,预测其编码一个由538个氨基酸组成的多肽,其N-端是一个由11个跨膜结构组成的疏水区域,C-端是一个亲水的尾巴。序列比对表明,NnNHX1与葡萄NHX1等的同源性较高,并且都具有高度保守的氨氯吡嗪咪结合位点序列LFFIYLLPPI。系统进化树表明NnNHX1与液胞膜型Na+/H+逆向转运蛋白关系较近,而与质膜型Na+/H+逆向转运蛋白关系较远。qRT-PCR结果显示NnNHX1基因在植株中属于组成型表达,但经NaCl诱导后,根中NnNHX1的表达量急剧增高后又迅速下降,而叶中的表达量却先缓慢增加后又剧烈下降。
The full-length cDNA sequence of the vacuolar Na + / H + antiporter gene NHX1 was isolated from lotus flowers by degenerate primers and RACE technique and named as NnNHX1. The gene is 2237bp in length and predicted to encode a polypeptide of 538 amino acids. Its N-terminal is a hydrophobic region consisting of 11 transmembrane structures and a C-terminal hydrophilic tail. Sequence alignment showed that NnNHX1 shared high homology with grape NHX1, and all had a highly conserved amino acid sequence LFFIYLLPPI. The phylogenetic tree indicated that NnNHX1 has a close relationship with the liquid membrane Na + / H + antiporter and a distant relationship with the plasma membrane Na + / H + antiporter. The results of qRT-PCR showed that NnNHX1 was constitutively expressed in plants. However, the expression of NnNHX1 in the roots was rapidly increased and then decreased rapidly after NaCl induction. However, the expression of NnNHX1 increased slowly and then dropped sharply.