Identification and characterization of a novel adenine phosphoribosyltransferase gene (ZmAPT2) from maize (Zea mays L.).

Suowei Wu, Zhanwang Yu, Fengge Wang, Weihua Li, Qingkai Yang, Chunjiang Ye, Yan Sun, Demin Jin, Jiuran Zhao, Bin Wang
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引用次数: 3

Abstract

Adenine phosphoribosyltransferase (APRT) is the key enzyme that converts adenine to adenosine monophosphate (AMP) in the purine salvage pathway. It was found that several different forms of APRT gene exist in plants, but no APRT gene in maize has been reported up to now. In this study, a novel maize APRT gene was cloned and characterized through a combination of bioinformatic, RT-PCR and RACE strategies. The full length of APRT cDNA sequence is 1202 nucleotides, with an ORF encoding 214 amino acid residues. Alignment of the deduced protein with that of other plant APRT genes indicates that the new gene is the form 2 of maize APRT, thus it was named ZmAPT2. Through basic local alignment search tool, search in the genomic survey sequence database of MaizeGDB, the putative genomic sequence of ZmAPT2 was obtained. Comparison of the cDNA and genomic sequence of the ZmAPT2 gene revealed that it contained seven exons and six introns. The locations of the introns within the maize ZmAPT2 coding region were consistent with those in the previously isolated APRTs of arabidopsis and rice. RT-PCR analysis showed that ZmAPRT was constitutively expressing in different organs under high temperature and salt stresses. Southern blot analysis indicated that at least three APRT genes existed in maize genome. These results confirmed that the novel maize ZmAPT2 gene was truly identified, and its potential role in maize growth and development was discussed.

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玉米腺嘌呤磷酸核糖基转移酶基因(ZmAPT2)的鉴定与鉴定
腺嘌呤磷酸核糖基转移酶(APRT)是嘌呤回收途径中将腺嘌呤转化为单磷酸腺苷(AMP)的关键酶。在植物中发现了几种不同形式的APRT基因,但目前还没有在玉米中发现APRT基因。本研究通过生物信息学、RT-PCR和RACE相结合的方法,克隆了一种新的玉米APRT基因,并对其进行了鉴定。APRT cDNA全长1202个核苷酸,ORF编码214个氨基酸残基。与其他植物APRT基因比对表明,该新基因为玉米APRT的2型基因,因此将其命名为ZmAPT2。通过基本的局部比对搜索工具,在MaizeGDB基因组调查序列数据库中进行搜索,得到ZmAPT2的推定基因组序列。比较ZmAPT2基因的cDNA和基因组序列发现,该基因含有7个外显子和6个内含子。内含子在玉米ZmAPT2编码区内的位置与先前分离的拟南芥和水稻APRTs中的位置一致。RT-PCR分析显示,高温和盐胁迫下,ZmAPRT在不同器官中均有组成性表达。Southern blot分析表明,玉米基因组中至少存在3个APRT基因。这些结果证实了ZmAPT2基因在玉米生长发育中的潜在作用。
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