GPAT基因表达载体的构建及对非洲菊的遗传转化

Construction plasmid about a cold resistant gene (GPAT) and transformation into Gerbera hybrida

运用已克隆的强抗冷植物兵豆的甘油-3-磷酸转酰酶(GPAT)基因构建表达载体,将GPAT和QS115质粒中的诱导型启动子串联后,插入带有终止子PinⅡ的表达载体pCAMBIA1300质粒中。并用PCR和酶切等多种方法进行鉴定。构建后的载体经PCR和酶切鉴定表明目的基因和启动子均已按预计方式插入载体的指定位置。用农杆菌介导的方法将它们转化到非洲菊中,获得了潮霉素的抗性植株,并通过PCR扩增验证,有抗性苗含有这个抗冷基因。

To construct a plasmid about a cold resistant gene （GPAT） from Lensculinaris medic for developing the genetical engineer, the plasmid was made as follows ：the ABA-responsive promoter was linked with GPAT,which come from pQS115 ,and then,they were inserted in the pCAMBIA1300. The pCAMBIA1300 contained Pin Ⅱ which come from pSBG700. PCR and restriction analysis were used to confirm the new plasmid. PCR and restriction analysis shewed that the plasmid pCAMBIA1300/GPAT was constructed successfully. It was transformated into Gerbera hybrida by agrobacterium. Hyg resistant transformants were obtained. One transgenic plant was obtained by PCR analysis,whlch indicated that GPAT gene was integrated into the plant.