【摘要】 通过Trizol法提取骆驼刺叶片总RNA的方法,反转录合成c DNA,PCR扩增得到一条600-800bp的DNA条带,与报道的Asp NHX1基因的核心序列基本相符,经3′-RACE扩增的DNA片段并连接到载体上进行测序.测序结果显示c DNA长度为1484bp,编码470个氨基酸,与猪毛菜Sa NHX1(Salsola affinis)、梭梭Ha NHX1(Haloxylon ammodendron)、盐爪爪Kf NHX1(Kalidium foliatum)的核苷酸序列同源性高达96%、89%、89%,氨基酸序列同源性达到97%、93%和90%.生物信息学分析显示As NHX蛋白的分子式为C4578H7674N1484O1907S291,蛋白的分子量为123347.5,理论等电点p I为5.01,不稳定参数为46.41,是一个富含疏水性氨基酸的不太稳定蛋白,理论上有35个苏氨酸的磷酸化位点,属于NHX1家族.说明己经在骆驼刺叶片中成功的克隆到Na+/H+逆向转运蛋白基因c DNA序列的部分片段,为下一步克隆骆驼刺叶片Na+/H+逆向转运蛋白基因的c DNA全序列提供了可能.更多还原

【Abstract】 Alhagi sparsifolia is a very good biological resistance material with the characteristics of cold resistance,drought resistance, salt resistance and wind characteristics. In order to study on the core sequence of the Alhagi sparsifolia’s NHX1 gene, Extracted the total RNA from the Alhagi sparsifolia’s leaves used Trizol kit method,synthesis the c DNA,and amplified by PCR,gained a DNA belt about 600 bp to 800 bp,which conforms to the core sequence of the NHX1 gene that had published.Amplified and sequenced the core As-NHX by 3′-RACE. The sequence result shows the length of the As-NHX is 1484 bp,encode 470 amino acid. The nucleotide sequence homology is 96%,89%and 89%,The amino acid sequence homology is 97%, 93% and 90% by compared the sequence with NHX1 between Sa NHX1(Salsola affinis),Ha NHX1(Haloxylon ammodendron)and Kf NHX1(Kalidium foliatum)reported in NCBI. The result analyses with bioinformatics shows the formula of As-NHX protein is C4578H7674N1484O1907S291,Mr is 123347.5,the isoelectric point(p I)is 5.01,it is a kind of unstable protein with rich hydrophobic amino acids,35 Threonine phosphorylation sites. We had cloned the fragment of NHX from Alhagi sparsifolia leaf and lay a foundation for the molecular biology research works of the resistance aspects on Alhagi sparsifolia.更多还原