拟南芥AtGPAT9基因cDNA的克隆及其转化大豆的研究

Cloning of AtGPAT9 cDNA from Arabidopsis and Its Transformation of Soybean

甘油-3-磷酸酰基转移酶(Glycerol-3-phosphate acyltransferases,GPATs;EC 2.3.1.15)是植物种子中储藏性油脂三脂酰甘油(Triacylglycerols,TAGs)合成中的一个关键酶,它催化甘油-3-磷酸分子sn-1位的酰化反应,形成溶血磷酯酸(Lysophosphatidic acid,LPA),从而起动TAGs的生物合成。拟南芥内质网中表达的AtGPAT9在此过程中可能起着重要作用。运用RT-PCR方法克隆到AtGPAT9基因的编码区cDNA,并构建了其植物表达载体,然后通过农杆菌介导的子叶节法对大豆进行了遗传转化,获得抗草丁膦(Phosphinothricin/PPT)筛选剂的再生植株。

Glycerol-3-phosphate acyltransferases（GPATs）play crucial role in the biosynthesis of triacylglycerols（TAGs）which are major form of the storage oils accumulated in developing seeds,and the enzymes catalyze the acylation of glycerol-3-phosphate at the sn-1 position and result in the formation of lysophosphatidic acid,which is the initial step of TAG biosynthesis.The endoplasmic reticulum-expressed AtGPAT9,a member of the GPAT family in Arabidopsis thaliana,was predicted to play an important role in the reaction.In the present study,the complete coding region cDNA of AtGPAT9 was cloned by using reverse transcriptase-mediated PCR,and the gene＇s plant expression vector was constructed.Then soybean transformation was performed by the Agrobacterium tumefaciens-mediated cotyledon node method,and the regenerated plants resistant to selective agent phosphinothricin/PPT were obtained.