Cloning for Full-length Pyruvate Orthophosphate Dikinase Gene of Sugarcane / 中国生物工程杂志

ABSTRACT

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 digest sites(XhoI and NotI) were introduced into primers,the full-length PPDK gene was splitted to two parts,and was ligated into pMD18-T simple vectors,respectively.Whole PPDK gene clone of sugarcane was finished.The strains were storaged in the lab.