猕猴桃L-艾杜糖脱氢酶基因的克隆及转化

Cloning and Transformation of L-Idonate Dehydrogenase Gene from Kiwifruit

以猕猴桃果实为材料,利用电子克隆技术获得了L-艾杜糖脱氢酶(L-Idonate dehydrogenase,IDH)基因,然后将其转入番茄中,为了解猕猴桃IDH基因如何调控AsA的降解奠定基础。结果表明,获得的猕猴桃IDH基因cDNA全长为1 239bp,含有一个1 080bp的开放阅读框,编码359个氨基酸;成功构建了IDH基因植物表达载体pWR-IDH及工程农杆菌,以番茄叶片及茎段为受体,通过农杆菌介导法进行转化,获得了4株经PCR和RT-PCR检测呈阳性的转基因植株。

For understanding degradation of ascorbic acid in plant,the L-Idonate dehydrogenase（IDH） gene was obtained from fruit of kiwifruit by in Silico cloning.The pWR-IDH vector was constructed and transformed into tomato.The results indicated that cDNA of IDH from kiwifruit is 1 239 bp and has an 1 080 bp open reading frame（ORF） and encodes a protein of 359 amino acid residues.The pWR-IDH vector was constructed and transformed into tomato leaves and shoot fragments by Agrobacterium tumefaciens-mediated transformation.Four transgenic resistant plants proved by PCR and RT-PCR suggested that the IDH gene had integrated into tomato genome.