转水稻巯基蛋白酶抑制剂基因毛白杨的获得

Obtaining the Transgenic Plants of Populus Tomentosa with Oryzacystatin Proteinase Inhibitor Insecticidal Gene

通过农杆菌介导法将水稻巯基蛋白酶抑制剂基因导入毛白杨。毛白杨叶片与农杆菌共培养， 采用卡那霉素３０ｍ ｇ／Ｌ筛选转化子获得抗性芽， 然后转到卡那霉素５０ｍ ｇ／Ｌ的培养基上继续筛选， 待转化芽长至１～２ｃｍ 时切下移至卡那霉素５０ｍ ｇ／Ｌ的生根培养基上获得完整再生植株。通过ＰＣＲ检测和ＰＣＲ－Ｓｏｕｔｈｅｒｎ

Oryzacystatin gene expressing a potential cysteine proteinase inhibitor against Coleoptera pests was introduced into Populus tomentosa by Agrobacterium mediated transformation. After cocultivation of leaf discs of P.tomentosa with Agrobacterium harbored Oryzacystatin gene, kanamycin resistance shoots were screened on selective medium with kanamycin 30mg/L and 50mg/L. The presence of Oryzacystatin gene in host cell the poplar genomes was confirmed by PCR and PCR Southern analysis.