农杆菌介导的大花蕙兰转ICE1基因

Transformation of ICE1 Gene into Cymbidium grandiftorium Mediated by Agrobacterium

大花蕙兰属热带植物,在北方组培成本很高,从拟南芥的RNA中克隆到耐寒基因(COR)的转录激活因子ICE1,连上CaMV35S启动子构建成植物表达载体pCAM BIA1300-35S-ICE1-poly,通过农杆菌介导的方法转化大花蕙兰。经过抗性筛选、诱导生芽、诱导生苗、诱导生根等过程,获得了70多棵潮霉素抗性植株,PCR鉴定12.9%为阳性,且部分PCR阳性植株通过了Southern检测,确定为转基因植株。

Cymbidium grandiftorium is tropical plant,the price of plant tissue culture is high in the northeast.This research cloned cold tolerance gene ICE1 which is a transcriptional active factor of cold tolerance gene（COR）from Arabidopsis＇s RNA,connected to CaMV35S and constructed into the plant expressing vector pCAMBIA1300-35S-ICE1-poly.Cymbidium grandiftorium was transformed by Agrobacterium Tumefaciens.This research got over 70 hygromycin resistant plants by resistance choice,inducing bud,inducing plant and inducing root.PCR identification showed that about 12.9% were positive and a part of PCR positive plants were positive by Southern analysis,finally these plants was determined for transgenic plants.