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Cloning and Sequence Analysis of pdk Gene in [Echinochloa crusgalli var. frumentacea (Roxb.) W. F. Wight]

家稗丙酮酸磷酸双激酶(PPDK)基因的克隆及序列分析


为揭示C4植物家稗[Echinochloa crusgalli var frumentacea(Roxb.)W.F.Wight] PPDK结构和功能特点,探索改善作物高光效途径,采用RT-PCR技术,克隆了家稗[Echinochloa crusgalli var frumentacea (Roxb.) W. F. Wight]丙酮酸磷酸双激酶(PPDK)基因的cDNA全长。核苷酸序列分析表明, 该片段全长3 085 bp, 包含一个2 844 bp的ORF,编码948个氨基酸(GenBank登录号:AY 792619)。编码区核苷酸序列与高粱(Sorghum bicolor)、甘蔗(Saccharum officinarum)、玉米(Zea mays)等C4pdk基因同源率分别为85.1%、84.0%和82.0%;与水稻[Oryza sativa (japonica cultivar-group)] pdk基因同源性也高达82.6%。氨基酸序列进化树分析表明其与玉米、水稻等禾本科植物最为接近;半定量RT-PCR研究表明,pdk基因仅在绿色叶片中表达。

Pyruvate orthophosphate dikinase(PPDK)plays an important role in C4 photosynthesis. In order to elucidate structural and functional characters of PPDK from [Echinochloa crusgalli var. frumentacea (Roxb.) W. F. Wight], and search a new approach to improve crop photosynthesis, a full-length cDNA for PPDK was isolated from [E. crusgalli var. frumentacea (Roxb.) W. F. Wight] by RT-PCR method. Sequencing analysis showed that the sequence length was 3 085 bp, including one ORF with 2 844 bp which coded a peotide of 948 amino acids. After alignment on ClustalW program, the identities of the cloned fragment with PPDK genes from C4 plants Sorghum bicolor, Saccharum officinarum and Zea mays were about 85.1%, 84.0%, and 82.0% respectively; and that from C3 plant [Oryza sativa (japonica cultivar-group)] was about 82.6%. The sequence has been submitted to the GenBank database, the accession number is AY792619. Alignment and phylogenetic analysis of the amino acid sequence deduced from the fragment and the PPDK sequences of other plants retrieved from GenBank were carried out by DNAstar program showing the sequence was close to Oryza sativa and Zea mays. Semi-quantitative RT-PCR analysis showed that PPDK was expressed exclusively in leaves.


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