【摘要】 目的:从普通小麦品种EM12中克隆ζ-胡萝卜素脱氢酶(ζ-carotene desaturase,ZDS)基因的cDNA全长序列。方法:根据已报道的EST序列,应用cDNA末端快速扩增(RACE)技术,对EST两端未知序列进行扩增、克隆和测序。结果:克隆得到cDNA全长序列2150bp,其ORF序列为1707bp,预测编码568个氨基酸残基,推测蛋白质分子量62.5kDa,带有一段19aa的信号肽序列。结论:根据同源序列分析,该序列与水稻ZDS蛋白的同源性为97%,与玉米ZDS蛋白的同源性为91%,而且与其他高等植物的ZDS都具有较高的同源性。该基因序列的分离与克隆为进一步开展小麦β-胡萝卜素生物合成机制的研究和利用转基因技术提高小麦β-胡萝卜素含量的研究奠定了基础。更多还原

【Abstract】 Objective:To isolate and sequence ζ-carotene desaturase (ZDS) cDNA from an elite wheat (Triticum aestivum L.) cultivar EM12. Method: According a reported EST sequence of wheat ZDS gene, rapid amplification of cDNA ends (RACE) approach was used to amplify full-length cDNA sequence. Result: The reconstructed cDNA sequence was 2 150bp in size, which contained a 1 707bp open reading frame (ORF) sequence. The predicted protein has a sequence of 568 amino acid residues with a molecular weight of 62.5kDa and a 19aa signal peptide sequence. Conclusion: Phylogenetic analysis demonstrated that ZDS obtained from wheat was 97% and 91% homologous with that from rice (Oryza sativa L.) and maize (Zea Mays L.) respectively at amino acid level, and it also showed high homology with other ZDS proteins. The results of the present study provided with the basis for further investigate the mechanism and genetic manipulation of β-carotene biosynthesis in wheat.更多还原