盐穗木Hc2a1基因的克隆及与表达分析  

Cloning and Expression Analysis of Salt- stress Responsive Hc2a1 Gene from Halostachys caspica

作　　者：苏立波[1] 艾力扎提.艾力 刘柳[1] 彭丹[1] 杨艺[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院/新疆生物资源与基因工程重点实验室,乌鲁木齐830046

出　　处：《新疆农业科学》2014年第9期1686-1691,共6页Xinjiang Agricultural Sciences

基　　金：新疆大学国家级大学生创新创业训练计划项目(201310755011)

摘　　要：【目的】盐胁迫能够抑制植物生长。探讨盐穗木(Halostachys caspica)盐胁迫响应基因表达变化的影响,有助于阐明盐穗木的耐盐分子机理。【方法】利用RACE方法获得盐穗木Hc2a1基因全长序列,并进行生物信息学分析和基因表达检测。【结果】Hc2a1基因开放阅读框为2 277 bp,编码758个氨基酸,蛋白质分子量为87.29 KDa,理论等电点(pI)为9.32。亚细胞定位于线粒体内膜,具有一个16个氨基酸的信号肽,含多个跨膜结构域,亲水性氨基酸残基多于疏水性氨基酸残基,二级结构多为无规则卷曲。利用荧光定量PCR检测显示,在高盐浓度600 mmol/L处理不同时间条件下,随着盐胁迫时间的延长,Hc2a1的表达量逐渐上升,12h达到最高峰,随后开始下降。【结论】盐穗木Hc2a1基因编码多个C2结构域和跨膜区的蛋白质,基因表达受盐胁迫的诱导。[Objective] Salt stress can inhibit the plant growth,which is helpful to explain the molecular mechanism of salt tolerance in halophytes to investigate the gene expression in response to salt stress [Method] The whole length sequence of Hc2a1 gene from Halostachys caspica was cloned By RACE （Rapid Amplification of cDNA Ends） and bioinformatics analysis and gene expression detection were carried out.[Result] The results showed that the open reading frame of Hc2a1 gene was 2 277 bp,encoding 758 amino acids with 87.29 KDa of molecular weight and 9.32 of theoretical pI.Hc2a1 protein was probably located in mitochondrial inner membrane and had a signal peptide of 16 amino acids and no transmembrane domain.A hydrophilic amino acid residue was more than hydrophobic amino acid residue.In the secondary structure,random coil was the majority of structural elements.Fluorescence quantitative PCR showed that the expression of Hc2a1 gene at different times under the high salt concentration of 600 mmol/L increased gradually and reached a peak at 12 h,then began to decline.[Conclusion] The results showed that the Hc2a1 gene from Halostachys caspica coding multiple C2 domains and transmembrane proteins,the expression of which was induced by salt stress.

关 键 词：盐穗木 Hc2a1基因 克隆 生物信息学分析 基因表达 

分 类 号：S188[农业科学—农业基础科学]