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海带lhcf4基因克隆和配子体差异表达研究

DOI: 10.7668/hbnxb.2011.06.009, PP. 41-49

Keywords: 海带,配子体,lhcf4,基因克隆,序列分析,差异表达

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Abstract:

根据糖海带(Laminariasaccharina)光捕获蛋白基因lhcf4的cDNA全长序列(GenBank登录号:AF226860)设计引物,通过RT-PCR方法获得海带(Saccharinajaponica)配子体lhcf4基因的cDNA全长序列,共1042bp,编码一个含218个氨基酸的前体蛋白lhcf4,推测分子量为23.11kDa,等电点为5.36。与其同源基因相似度分析表明,lhcf基因的开放阅读框(Openreadingframe,ORF)在家族内及物种间高度保守,变异主要发生在非翻译区(Untranslatedre-gion,UTR)。预测lhcf4成熟蛋白三级结构存在4个α螺旋区,其中第一、三螺旋较长,形成跨膜螺旋,第二个螺旋相对较短,完全镶嵌在膜中,另外一个为短螺旋,位于类囊体膜-类囊体腔界面上。杂色藻类、绿藻及豌豆光捕获蛋白氨基酸多序列比对显示叶绿素a结合位点及盐桥形成氨基酸残基在植物进化中高度保守;将预测的海带lhcf4蛋白3D结构与豌豆X-衍射晶体结构进行空间重叠,获得了结合在lhcf4蛋白上的5个叶绿素a的空间分布。Neighbor-joining分子进化树显示原绿球藻、绿藻和杂色藻类的光捕获蛋白基因是三个独立进化的系统,共同起源于蓝细菌的只结合叶绿素a的光捕获蛋白;海带lhcf4基因与糖海带lhcf4及巨藻的fcpb基因为直系同源;糖海带lhcf4、lhcf7及lhcF1基因为旁系同源。实时荧光定量PCR结果证实lhcf4基因具有雌、雄配子体差异表达特征。

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