Wu Suowei, Yu Zhanwang, Wang Fengge, Li Weihua, Yang Qingkai, Ye Chunjiang, Sun Yan, Jin Demin, Zhao Jiuran, Wang Bin
The State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, People's Republic of China.
DNA Seq. 2008 Jun;19(3):357-65. doi: 10.1080/10425170701606235.
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.
腺嘌呤磷酸核糖转移酶(APRT)是嘌呤补救途径中催化腺嘌呤转化为单磷酸腺苷(AMP)的关键酶。研究发现,植物中存在几种不同形式的APRT基因,但截至目前尚未见玉米APRT基因的报道。本研究通过生物信息学、RT-PCR和RACE技术相结合,克隆并鉴定了一个新的玉米APRT基因。该基因cDNA全长1202个核苷酸,开放阅读框编码214个氨基酸残基。将推导的蛋白质序列与其他植物APRT基因进行比对,表明该新基因是玉米APRT的2型,因此命名为ZmAPT2。通过基本局部比对搜索工具(BLAST)在MaizeGDB的基因组Survey序列数据库中搜索,获得了ZmAPT2的推定基因组序列。ZmAPT2基因的cDNA和基因组序列比较显示,该基因含有7个外显子和6个内含子。玉米ZmAPT2编码区内内含子的位置与拟南芥和水稻中先前分离的APRTs一致。RT-PCR分析表明,ZmAPRT在高温和盐胁迫下的不同器官中组成型表达。Southern杂交分析表明,玉米基因组中至少存在3个APRT基因。这些结果证实了新的玉米ZmAPT2基因的真实身份,并对其在玉米生长发育中的潜在作用进行了讨论。