王子剑, 孙丹, 于颖, 张岩, 王超. 白桦扩展蛋白基因的克隆分析及植物表达载体构建[J]. 西南林业大学学报, 2016, 36(1): 1-8. DOI: 10.11929/j.issn.2095-1914.2016.01.001
引用本文: 王子剑, 孙丹, 于颖, 张岩, 王超. 白桦扩展蛋白基因的克隆分析及植物表达载体构建[J]. 西南林业大学学报, 2016, 36(1): 1-8. DOI: 10.11929/j.issn.2095-1914.2016.01.001
Wang Zijian, Sun Dan, Yu Ying, Zhang Yan, Wang Chao. Gene Cloning and Construction of Plant Expression Vector of Expansin Genes in Betula platyphylla[J]. Journal of Southwest Forestry University, 2016, 36(1): 1-8. DOI: 10.11929/j.issn.2095-1914.2016.01.001
Citation: Wang Zijian, Sun Dan, Yu Ying, Zhang Yan, Wang Chao. Gene Cloning and Construction of Plant Expression Vector of Expansin Genes in Betula platyphylla[J]. Journal of Southwest Forestry University, 2016, 36(1): 1-8. DOI: 10.11929/j.issn.2095-1914.2016.01.001

白桦扩展蛋白基因的克隆分析及植物表达载体构建

Gene Cloning and Construction of Plant Expression Vector of Expansin Genes in Betula platyphylla

  • 摘要: 从白桦中分离了4条扩展蛋白家族基因全长cDNA序列和2条大于400bp的部分cDNA序列,命名为BpEXP1~BpEXP5和BpEXPL,其中4个全长cDNA推导蛋白氨基酸残基数为186~258个,编码蛋白的分子量为2007~2780kDa,理论等电点为557~920。氨基酸序列分析结果表明:5个alpha- expansin序列有共同的结构特征,即在N-末端有8个保守的半胱氨酸残基的丰富域,C-末端有4个保守的色氨酸残基的丰富域,中间有1个组氨酸功能域。利用Real time RT-PCR分析EXP家族基因在白桦不同器官和组织内的表达模式,结果表明:6个EXP基因在白桦的不同器官和组织中有着不同的表达水平,其中BpEXP1在分生木质部中表达量最高,推测其与木材形成过程的细胞壁修饰相关;BpEXP3在发育的蒴果中表达量最高,推测其与果实和种子发育过程中的细胞壁修饰相关。最后选择BpEXP1和BpEXPL基因,构建了pROKⅡBpEXP1和pROKⅡBpEXPL植物表达载体。

     

    Abstract: Four fulllength cDNAs and two partial cDNAs, designated BpEXP1 to BpEXP5, and BpEXPL, encoding expansin homologues, were isolated from Betula platyphylla . The open reading frames of four fulllength cDNAs ranged from186 to 258 amino acids, with predicted molecular weights of 242 to 258 kDa and PI of 557 to 920.The deduced amino acid sequences of the five alphaexpansin include eight typical Cys residues in the N-termini of the proteins, four Trp residues near the C-termini and an HFD motif in the central region, which are all characteristic of expansins. Real time RT-PCR analyses in birch organs and tissues showed that BpEXP1 was specifically expressed in developing xylem exhibiting secondary growth, and was presumed that BpEXP1 was involved in the cell wall modification during the wood formation. By contrast, BpEXP3 was mostly expressed in developing capsule, and then was presumed that BpEXP3 was involved in the cell wall modification during the seed development. Finally, BpEXP1 and BpEXPL were chosen to construct the plant expression vector of pROKII-BpEXP1 and pROKII-BpEXPL.

     

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