甘蔗全长丙酮酸磷酸二激酶(PPDK)基因的克隆  

Cloning for Full-length Pyruvate Orthophosphate Dikinase Gene of Sugarcane

作　　者：杨福明[1] 徐德昌[1] 侯爱菊[1] 

机构地区：[1]哈尔滨工业大学食品科学与工程学院,哈尔滨150090

出　　处：《中国生物工程杂志》2007年第10期87-92,共6页China Biotechnology

基　　金：黑龙江省科技攻关资助项目(GB05B104)

摘　　要：和C3植物相比,C4植物具有明显的生长优势及水分和营养利用率,生物产量也较高。甘蔗是典型的C4作物之一。以甘蔗叶片提取的基因组DNA为模板,以GenBank公布的甘蔗PPDK基因cDNA序列设计引物,进行LA-PCR(Long Acute PCR)扩增。将PCR产物克隆到pMD18-T载体中,转化大肠杆菌JM109,测序,得到了13.5kb的甘蔗全长PPDK基因序列。为方便后续实验,在引物中引入可利用的XhoI和NotI酶切位点,将全长PPDK基因分两段克隆到pMD18-TSimple载体中,转化大肠杆菌JM109,完成了甘蔗全长丙酮酸磷酸二激酶(PPDK)基因的完整克隆,为将其导入C3作物中奠定了研究基础。Compared with C3 plant, C4 plant had evident growth advantage, higher rates of water and nutrition using, and higher bio-yield than C3 plant, Sugarcane was one of the typical C4 plants. DNA was extracted from sugarcane leaves, primers were designed by the cDNA sequence of PPDK gene from GenBank. Then DNA was amplified by LA-PCR （Long Acute PCR） method, ligated into pMD18-T vectors, and transformed into E. coli. JM109. Full-length PPDK gene sequence of sugarcane was obtained, the sequence was 13.5kb in length. For convenient of the next experiment, two full-length PPDK gene was splitted to two parts, digest sites（ XhoI and and was ligated into NotI） were introduced into primers, pMD18-T simple vectors, respectively. Whole PPDK gene clone of sugarcane was finished. The strains were storaged in the lab.

关 键 词：甘蔗 PPDK基因 LA-PCR技术 全长DNA克隆 

分 类 号：Q78[生物学—分子生物学]