川芎甜菜碱醛脱氢酶cDNA的克隆及其植物表达载体的构建  

Cloning of Betaine Aldehyde Dehydrogenase cDNA in Liqusticum chuanxiong and Construction of its Plant Expression Vector

作　　者：毛莹[1] 张艳军[1] 王万军[1] 陈劲松[2] 廖海[2] 周嘉裕[2] 

机构地区：[1]西南交通大学生命科学与工程学院,四川成都610031 [2]中国科学院成都生物研究所,四川成都610041

出　　处：《北方园艺》2013年第5期87-90,共4页Northern Horticulture

基　　金：国家自然科学基金资助项目(31271302);教育部中央高校基本科研业务费专项资金资助项目(SWJTU11CX114);中国科学院成都生物研究所山地生态恢复与生物资源利用重点实验室及生态恢复与生物多样性保育四川省重点实验室开放课题资助项目;国家级大学生创新创业训练计划资助项目(201210613050);西南交通大学科技发展计划资助项目(2008B06)

摘　　要：以川芎为试材,从叶片中提取总RNA,经过RT-PCR获得甜菜碱醛脱氢酶(Betaine aldehyde dehydrogenase)基因的cDNA,纯化后与pMD18-T载体连接,转化大肠杆菌Top10,获得甜菜碱醛脱氢酶全长基因序列,以期构建植物表达载体。结果表明:甜菜碱醛脱氢酶全长核苷酸长度为1 527bp,编码508个氨基酸。与GenBank中已发表序列HM35276进行比较,核苷酸同源性为100%。将该基因片段克隆到植物表达载体pBI121中,构建重组质粒pBI121/Betaine alde-hyde dehydrogenase,并将所获重组质粒经过双酶切和PCR处理后进行序列测定,证实表达载体上含有目的片段,且连接、构建正确,为BADH的进一步表达奠定了基础。Taking Liqusticurn chuanxiong as material, the total RNA was extracted from leaf of Liqusticum chuanxiong and cDNA of Betaine aldehyde dehydrogenase gene were obtained using RT-PCR. The purified RT-PCR products were constructed into pMD18-T vector. The results showed that the full length of BADH gene consists of 1 527 bp, which encoded 508 amino acids. The homology of its nucleotide sequence with HM35276 was 100%. Furthermore,the aim gene was cloned into plant expression vector pBI121 and a recombinant plasmid pBI121/Betaine aldehyde dehydrogenase were constructed successfully according to double enzyme digestion,PCR and sequencing.

关 键 词：川芎 甜菜碱醛脱氢酶 克隆 植物表达载体 

分 类 号：S567.239[农业科学—中草药栽培]