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水稻OsPP6C基因的克隆、过表达载体构建与生物信息学分析

DOI: 10.7668/hbnxb.2013.05.011, PP. 59-65

Keywords: 水稻,蛋白磷酸酶6,过表达载体构建,生物信息学分析

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Abstract:

为克隆水稻蛋白磷酸酶6(PP6)催化亚基基因OsPP6C,并构建该基因的过表达载体以及进行生物信息学分析。采用RT-PCR技术从水稻中花11叶片cDNA中扩增OsPP6C基因全长,并构建与GUS融合的pBI121-OsPP6CGUS表达载体。通过生物信息学方法分析了OsPP6C蛋白的理化性质、与拟南芥AtPP6C(即AtFyPP)之间的同源性以及不同物种间的系统发育关系,此外,对OsPP6C基因的启动子进行了顺式作用元件分析。结果表明,该基因转录序列包含一个912bp的开放阅读框,编码303个氨基酸残基,蛋白的分子量约为3.47kDa,等电点为5.13;具有疏水性,但不存在信号肽,属于非分泌型蛋白。OsPP6C与拟南芥AtFyPP1和AtFyPP3氨基酸序列一致性分别为93.73%和93.40%,系统进化树显示OsPP6C与小麦TaPP6C的亲缘关系最近,而与其他物种亲缘关系相对较远。对该基因启动子中含有的顺式作用元件分析表明,该启动子中除TATA盒和CAAT盒外,还含有多种参与光应答、激素(ABA、乙烯、生长素、MeJA和赤霉素等)应答以及低温、热激和干旱等非生物胁迫因子应答的调控元件。为进一步研究水稻OsPP6C基因的表达特性和功能奠定了基础。

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