苹果脱氢抗坏血酸还原酶(DHAR)基因cDNA片段的克隆及序列分析

Cloning and sequence analysis of cDNA segment encoding dehydroascorbate reductase from malus

以皇家嘎拉苹果叶片为材料提取总RNA,合成cDNA第一链后,先利用巢式PCR获得苹果脱氢抗坏血酸还原酶(DHAR)基因中间片段,再利用3-′RACE方法进一步获得3′末端核酸序列,共得到840 bp包含完整3′末端的DHAR基因cDNA片段,其序列与烟草、水稻、番茄中DHAR基因序列的同源性均大于70%,证明所克隆到的核苷酸片段确为苹果DHAR基因cDNA片段。

In this study,a cDNA fragment of dehydroascorbate reductase （DHAR） including whole 3＇- end was cloned from Royal Gala by RT-（nest） PCR and 3＇-RACE. As deduced from the 840 bp cDNA nucleotide,it presented more than 70% homologous traits with tobacco DHAR,Rice DHAR and tomato DHAR,These results suggest that the cDNA fragment is from DHAR gene of Royal Gala.