摘要
为构建猪传染性胃肠炎病毒(TGEV)S基因转玉米植株,本研究以玉米自交系"丹598"和"H99"的胚性愈伤组织为材料,通过愈伤组织对卡那霉素的敏感性试验,确定了卡那霉素40mg/L～56mg/L为愈伤组织适宜选择压。利用农杆菌介导法将TGEV S基因导入玉米自交系中,并对农杆菌转化系统的条件进行了优化。结果表明,根癌农杆菌EHA105的菌液OD600nm值为0.5～0.6时,侵染20min为农杆菌转化的最适条件。并对转化的愈伤组织进行分化诱导出苗后进行PCR和Southern blot检测,证明外源目的基因已整合到玉米基因组中。
Transgenic maizes expressing transmissible gastro-enteritis virus（TGEV） S gene were established using embryogenic callus of maize inbred lines dan 598 and H99 as receptor materials.The TGEV S gene was cloned into an Agrobacterium binary vector and transformed into maize inbred lines by Agrobacterium under the selection of 40 to 56 mg/L of kanamycin.The factors influencing maize callus transformation were investigated,and optimal transformation was achieved using Agrobacterium tumefaciens EHA105 at concentration of OD600nm 0.5 to 0.6 and infection time of 20 min.The integration of TGEV S gene in the maize genome was confirmed by PCR and Southern blot analysis.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第10期752-756,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
吉林省科技厅资助项目(20080547)
关键词
猪传染性胃肠炎病毒
S基因
玉米
农杆菌
transmissible gastro-enteritis virus
S gene
maize
Agrobacterium
