玉米丝黑穗病菌与玉米互作中弹性蛋白的诱导表达与克隆
首发时间:2013-02-06
摘要:本实验室前期选用对丝黑穗病表现明显抗性差异的自交系Mo17(对丝黑穗病高抗)和黄早四(HZ4,对丝黑穗病高感)为试材, 采用SSH和cDNA芯片的方法,对两自交系的幼苗期接种与不接种处理,将两个材料的处理及对照mRNA分别与Mo17全生育期cDNA芯片杂交,对两种状态下所表达的差异基因进行比较分析,发现了一个诱导表达的EST序列,该基因在抗感自交系中表达差异两倍以上。本研究以前期筛选到的EST为基础,首先利用荧光定量PCR进行分析验证其在抗感自交系中的表达情况,然后采用同源序列克隆技术在Mo17和HZ4分离出这个基因, 并命名为SRR1。将获得的全长cds序列在NCBI数据库同源性比对,发现SRR1蛋白与玉米节肢弹性蛋白前体(pro-resilin precursor)同源性达到99%。B73、Mo17和HZ4自交系中,SRR1开放阅读框分别为1134 bp、1134bp和1125bp, 分别编码377、377和374个氨基酸。将B73、Mo17和HZ4的SRR1基因的cDNA序列和编码的氨基酸序列进行多序列同源比对,发现B73和Mo17的序列完全相同,HZ4和B73、Mo17有20处碱基发生了突变,编码的氨基酸有6处发生了变化。本研究构建了SRR1与GFP融合载体,洋葱表皮的瞬时表达结果表明,SRR1蛋白定位于细胞壁。
关键词: 玉米 玉米丝黑穗病 节肢弹性蛋白前体基因
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Cloning and expression of pro-resilin precursor gene association to interaction between maize and head smut fungus
Abstract:Head smut is one of important diseases of maize, which can cause great loss of corn yield in the world wide. Developing corn resistance germplasm and breeding for disease resistance is a fundamental way to control head smut. A pro-resilin precursor gene (named SRR1) was cloned in the inbred lines Mo17 and HZ4 using homologous sequences cloning method. The bioinformatic analysis showed open reading frame of the gene is 1134 bp、1134bp and 1125bp in B73、Mo17and HZ4,respectively. SRR1's ORF and GFP fusion protein was constructed. Onion epidermis transient expression results showed that the protein coded by the gene was located in the cell wall of maize.
Keywords: maize head smut pro-resilin precursor gene
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