新疆盐生植物猪毛菜逆向运输蛋白基因SaNHX1 3’-RACE的克隆及序列分析

以新疆盐生植物猪毛菜(Salsola collina Pall.)为材料提取总RNA,根据NHX1家族同源序列保守区设计1对简并引物,进行RT-PCR扩增,得到猪毛菜逆向运输蛋白基因cDNA中间一段619 bp序列。再用快速扩增cDNA 3’末端(3’RACE)方法,得到约1 000 bp 3’末端序列。将两段序列进行拼接,得到猪毛菜长为1 585bp(Gen Bank登陆号为EU072932)的逆向运输蛋白基因(SaNHX1)cDNA序列,编码370个氨基酸,其编码序列、氨基酸序列与同科耐盐植物盐角草相应序列同源性为85%和87%,研究为进一步克隆猪毛菜NHX1全基因序列和研究其功能打下基础。

Clone and Sequence Analysis of 3'RACE of Antiporter Protein Gene SaNHX1 in Xinjiang Halophytes Plant Salsola collina Pall.

Total RNA in this experiment was extracted from Xinjiang halophytes plant Salsola affinis(Salsola collina Pall.).According to NHX1 family homologous sequence conservative region,.one pair of degenerate primers was used for RT-PCR amplification.The results showed that a partial middle 619 bp fragment of SaNHX1 antiporter protein was obtained.The 3' end sequence was obtained by 3' RACE method.These two fragments were overlapped,getting SaNHX1 3'terminal fragment 1 585 bp sequence encoding 370 amino acid residues.The codon sequence and amino acid sequence of the 3'ends of SaNHX1 gene were shared by 85% and 87% identity with those of the same family halophytes SeNHX1 gene respectively.This research provided a basic foundation to clone the whole NHX1 sequence and study its structure and function.