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费尔干猪毛菜病程相关蛋白基因(SfPR-1)的克隆及在盐胁迫下的表达分析

Cloning of Pathogen-Related Protein Gene(SfPR-1) from Salsola ferganica and Its Expression Analysis under Salt Stress

  • 摘要: 利用抑制消减杂交法从藜科猪毛菜属盐生植物费尔干猪毛菜(Salsola ferganica)中分离得到了一个盐胁迫响应的cDNA片段,结合SMARTTM RACE技术获得了费尔干猪毛菜病程相关蛋白基因的cDNA,命名该基因为 SfPR-1 (GenBank登录号: JQ670917)。序列分析表明,SfPR-1长817 bp,含有501 bp的阅读框、 65 bp的 5'-UTR 和251 bp的3'-UTR,编码166个氨基酸,分子质量为18.01 kD,理论等电点为9.37。通过BLAST同源序列比对分析,结果显示该基因编码的蛋白与已知甜菜、拟南芥、烟草及玉米的病程相关蛋白PR-1同源性分别为73.6%、57.8%、55.5%和53.9%,且具有PR-1家族特有的6个半胱氨酸保守结构域。半定量RT-PCR和实时荧光定量RT-qPCR分析表明,该基因在盐胁迫后表达呈明显上调,初步推测病程相关蛋白基因SfPR-1可能与费尔干猪毛菜的耐盐性相关。

     

    Abstract: A cDNA fragment was isolated from Salsola ferganica by suppression subtractive hybridization and its full-length cDNA with 817 bp was cloned by SMARTTM RACE, and was named SfPR-1 gene (GenBank accession number: JQ670917). The SfPR-1 gene consisted of a 501 bp open reading frame encoding 166 amino acids with molecular weight of 18.01 kDa and an isoelectric point of 9.37, a 65 bp 5'-UTR and 251 bp 3'-UTR. The deduced amino acid sequence of SfPR-1 showed high identity of 73.6%, 57.8%, 55.5% and 53.9% to those of pathogen-related protein 1 from Beta vulgaris, Arabidopsis thaliana, Nicotiana tabacum and Zea mays, respectively, and had a conserved six-cysteine motif. Reverse transcriptase PCR and Real-time PCR methods were used to investigate the expression profile of the SfPR-1 gene under salt stress. SfPR-1 showed up-regulated expression patterns under salt treatment. Based on our results, we concluded that the SfPR-1 gene might be involved in salt response and an important component for the salt tolerant pathway in Salsola ferganica.

     

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