摘要
利用根癌农杆菌遗传转化系统，以普通小麦扬麦158为材料，将玉米泛生素基因启动子（Ubi）驱动下的磷酸烯醇式丙酮酸羧化酶（PEP Case）基因，以及潮霉素磷酸转移酶基因（hpt）导入预培养10～12d小麦胚性愈伤组织中。经过在含100-150mg·L^-1潮霉素（hyg）的筛选培养基上连续筛选，获得了hyg抗性植株。经GUS组织化学染色检测和PCR鉴定，其中的9棵含有Ecppc基因，PCR阳性植株比例为3．03％。初步鉴定已成功地建立了小麦遗传转化系统，为进一步探讨PEP Case基因在植物中的转化和表达奠定了基础。
PEPCase gene drived by ubiquiten promoter and hygromycin phosphotransferase gene hpt was transformed to yangmai158 mediated by Agrobacterium tumefaciens. Regeneration plant was inducted from 10- 12 day wheat immature embryos. Hygromycin resistant （hyg） plantlets were obtained by 2- 3 cycles screening in selective medium containing 100-150 mg·L^-1 hygomycin. By means of PCR amplification and GUS histochemical analysis of hyg-resistant plants showed that the target gene had been integrated into wheat accompany with hpt gene. The frequency of transformation was 3.03%.
出处
《山西农业大学学报（自然科学版）》
CAS
2009年第4期323-326,共4页
Journal of Shanxi Agricultural University(Natural Science Edition)
基金
山西省科技攻关项目(2007031027-3)
山西农业大学科技创新基金(2008027)
