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一个棉花微管结合蛋白基因GhSP1L5的克隆及表达分析

, PP. 82-87

Keywords: 棉花纤维,Spiral1-Like5基因,微管结合蛋白,表达

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Abstract:

利用RT-PCR技术从处于快速伸长发育时期的棉花纤维组织中克隆得到Spiral1-Like(SP1L5)基因(GhSP1L5),其开放读码框为315bp,编码105个氨基酸的蛋白质,生物信息学分析表明,GhSP1L5蛋白N端和C端较为保守,并含有SCOP结构域,是典型的微管结合蛋白。进化树分析表明该基因与SP1L5家族基因的亲缘关系较近。构建原核表达载体pET28a-GhSP1L5,转化大肠杆菌BL21(DE3)并进行体外诱导表达,通过SDS-PAGE检测目的蛋白的表达,获得分子质量约为12kD的重组蛋白。QRT-PCR结果分析表明GhSP1L5基因在开花后15和20dpa的纤维组织中表达量较高,在棉纤维发育的其他时期及根、茎、叶组织中表达量较低。

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