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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.
| Abbreviations | ||
| APRT | = | adenine phosphoribosyltransferase |
| AMP | = | adenosine monophosphate |
| PRPP | = | 5-phosphoribosyl-1-pyrophosphate |
| NCBI | = | National Center for Biotechnology Information |
| MaizeGDB | = | Maize Genetics and Genomics Database |
| BLAST | = | basic local alignment search tool |
| RACE | = | rapid amplification of cDNA ends |
| RT-PCR | = | reverse transcription polymerase chain reaction |
| EST | = | expressed sequence tag |
| GSS | = | genomic survey sequence |
| ORF | = | open reading frame |
| GSP | = | gene specific primers |
| NGSP | = | nested gene specific primers |
| TGMS | = | thermo-sensitive genic male sterility |
| Abbreviations | ||
| APRT | = | adenine phosphoribosyltransferase |
| AMP | = | adenosine monophosphate |
| PRPP | = | 5-phosphoribosyl-1-pyrophosphate |
| NCBI | = | National Center for Biotechnology Information |
| MaizeGDB | = | Maize Genetics and Genomics Database |
| BLAST | = | basic local alignment search tool |
| RACE | = | rapid amplification of cDNA ends |
| RT-PCR | = | reverse transcription polymerase chain reaction |
| EST | = | expressed sequence tag |
| GSS | = | genomic survey sequence |
| ORF | = | open reading frame |
| GSP | = | gene specific primers |
| NGSP | = | nested gene specific primers |
| TGMS | = | thermo-sensitive genic male sterility |
Acknowledgements
This study was supported by Agricultural Innovative Platform Project of Beijing Natural Science Fund Program (Grant No.YZPT-02-06) and the State Key Basic Research and Development Plan of China (Grant No. 2006CB101700).