| 摘要: |
| L-半乳糖脱氢酶（L-Galactose dehydrogenase,GalDH）是维生素C合成的L-半乳糖途径中,催化L-半乳糖生成L-半乳糖内酯的关键酶.根据GenBank中登录的的GalDH cDNA序列设计1对扩增引物,以嘎拉苹果叶片为材料,采用RT-PCR法扩增出GalDH cDNA全长.将获得的基因片段克隆到pMD18-T载体上,转入大肠杆菌DH5α筛选阳性克隆,经酶切和PCR鉴定,并对插入片段进行序列分析,结果表明,本试验获得的cDNA片段长为1 111 bp,是苹果GalDH cDNA全长. |
| 关键词: 苹果 克隆 序列分析 |
| DOI： |
| 分类号: |
| 基金项目:西北农林科技大学“拔尖人才支持计划”项目 |
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| Cloning and sequence analysis of full-length cDNA encoding L-Galactose dehydrogenase from apple |
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| Abstract: |
| L-Galactose dehydrogenase (L-GalDH) is the key enzyme of ascorbate biosynthesis and oxidizing L-Gal to L-galactono-1,4-lactone.Full-length cDNA encoding L-GalDH was cloned from Royal Gala apple(Malus Pumila Mill cv.Royal Gala) leaves by RT-PCR method according to the homologous gene in GenBank. Then the target fragment was purified from agarose gels and integrated into pMD18T cloning vector.After being transformed into E.coli.DH 5a.tested by restriction endonuclease digestion and PC,the screened positive clone was sequenced. Sequencing analysis shows that the length of the cDNA of apple GalDH is 1 111 bp,which is the full-length cDNA of GalDH. |
| Key words: apple L-Galactose dehydrogenase RT-PCR cloning sequencing analysis |
