【摘要】 为了获得与盐碱胁迫相关的14-3-3蛋白基因的表达模式,基于羊草14-3-3蛋白的EST序列,通过RACE方法从羊草中克隆得到的Lc14-3-3基因,并利用生物信息学分析获得了其基因信息,利用荧光定量PCR（qRT-PCR）分析获得了羊草植株在Na₂CO₃胁迫下Lc14-3-3基因的表达模式。结果表明:Lc14-3-3基因编码区全长为786 bp,编码261个氨基酸,推测蛋白质分子量为29.26 k D,理论等电点（p I）为4.83,与已知的单子叶植物来源的同类基因同源性较高,相似性为88%¹00%。q RT-PCR表达模式分析表明,羊草14-3-3基因在无胁迫处理情况下有本底水平表达。在200 mmol/L的Na₂CO₃胁迫不同时间后,羊草叶片中Lc14-3-3基因的表达量在6²4 h之间表达量明显上升,但胁迫持续48 h时,Lc14-3-3表达量降至较低水平。但随着胁迫时间增加Lc14-3-3表达量骤增,96 h后高于对照组10倍左右。羊草根部的Lc14-3-3基因表达量在胁迫处理后6²4 h间为无胁迫条件的2倍,在胁迫处理48 h后下降,随着胁迫时间增加,96 h表达量为对照组11倍左右。此结果为进一步研究抗盐碱基因提供理论基础。更多还原

【Abstract】 In order to obtain the expression pattern of the 14-3-3 protein gene associated with saline-alkaline stress. Based on the 14-3-3 protein EST sequence of Leymus chinensis, a Lc14-3-3 gene was cloned from L.chinensis by RACE. And the genetic information was obtained by bioinformatics analysis, the expression pattern of Lc14-3-3 gene in Leymus chinensis plants under Na₂CO₃ stress was obtained by fluorescence quantitative PCR（q RT-PCR）. The results indicated that the full length of the coding region of Lc14-3-3 gene was 786 bp, encoding261 amino acids. The molecular weight of the protein was 29.26 k D and theoretical p I was 4.83. Homology analysis showed that the deduced amino acid sequence of Lc14-3-3 shared 88% ¹00% homology with 14-3-3from monocotyledon. q RT-PCR expression pattern analysis showed that the 14-3-3 gene of Leymus chinensis was expressed at the background level without stress. With the treatment of 200 mmol/L Na₂CO₃ at different time, the expression of Lc14-3-3 increased between 6²4 h in Leymus chinensis leaves, and then dropped after 48 h, the expression of Lc14-3-3 was reduced to a low level. With the extension of Na₂CO₃ stress time, the expression of Lc14-3-3 was increased dramatically. After treated for 96 hours showed ten times peak compared with control.The results of q RT-PCR analysis showed that the expression level of Lc14-3-3 gene in Leymus chinensis root treated for 6²4 h after 200 mmol/L Na₂CO₃ was two times compared with control. The expression of Lc14-3-3decreased to a low level after treatment for 48 h. As the increasing of stress time, the expression group at 96 h was11 times higher than that of the control group. The results provided a theoretical basis for the further study of saline-alkaline genes.更多还原