摘要:
采用同源克隆RT-PCR结合RACE技术,从福建省地方梨种质资源莆田豆梨中克隆出GPX基因的cDNA全长序列分离克隆并运用生物信息学方法对序列进行分析。克隆得到福建省地方种质资源莆田豆梨GPX基因932bp的cDNA全长序列,分析表明,该序列的5′端和3′端的非编码序列长度分别为204bp和221bp,开放阅读框为507bp,编码168个氨基酸。氨基酸序列与苹果、柑橘、龙眼、荔枝的同源性90%以上。GPX基因在GenBank上登录,登录号为JQ011278。生物信息学分析表明:GPX蛋白的分子量是20 083.0Da,理论等电点pI 5.61,是不具跨膜结构域的亲水性胞质蛋白,不具有信号肽,细胞主要定位在细胞质上,有1个区域最有可能形成卷曲螺旋,由26.58%的a螺旋,19.26%的延伸链和54.16%的不规则卷曲组成,磷酸化位点有12个。此外,还对GPX酶分子三维立体结构等进行预测和分析。
Abstract:
Aglutathione peroxidasegene(GPX)was cloned from the leaves of Putian pear,PyruscalleryanaDecne.The RT-PCR and RACE were applied to obtain the complete cDNA sequence for GPX.The full length of the cDNA was approximately 932 bp consisting of an open reading frame of 507 bpwith the 5′-and 3′-untranslated regions of204 bp and 221 bp,respectively.The putative protein had 168 amino acids and was greater than 90% homogenous on the sequence with Maluspumila,Citrus reticulata Banco,Dimocarpuslongan and Litchi chinensis.The gene had been registered in GenBankwith a code of JQ011278.The nucleotide and amino acid sequences of GPX were analyzed by using a bioinformatics software.The results showed that the protein had a molecular weight of 20083.0Da anda theoretical pI 5.61.It was a hydrophilic cytoplasmic protein without transmembrane domain and signal peptide.The cell was mainly located in the cytoplasm at a region most likely to form coiled-coilcontaining26.58% α-spiral,19.26%extending chain and 54.16%irregular curl.And,there were 12 phosphorylation sites on the cell.A3-Dmolecular structure of the enzyme was proposed with relevant analysis.
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