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农杆菌介导的河北杨遗传转化体系的建立
引用本文:王沛雅,杨晖,杨涛,张军,郭琪,强维亚,周剑平.农杆菌介导的河北杨遗传转化体系的建立[J].生物技术通报,2012,0(3):141-147.
作者姓名:王沛雅  杨晖  杨涛  张军  郭琪  强维亚  周剑平
作者单位:1. 甘肃省科学院生物研究所,兰州730000;甘肃省工业微生物工程技术研究中心,兰州730000
2. 兰州大学生命科学学院,兰州,730000
3. 甘肃省工业微生物工程技术研究中心,兰州730000;甘肃省科学院,兰州730000
基金项目:甘肃省国际科技合作项目,甘肃省科学院应用技术研究与开发计划项目,甘肃省工业生物过程工程创新服务平台资助项目
摘    要:以兼具生态和能源植物功能的木本模式植物——杨树(河北杨)为材料,研究了携带促生长基因(35S-DAS5)的根癌农杆菌载体介导的河北杨遗传转化若干因素对转化效果的影响。结果显示,较适宜的转化系统为预培养2-4 d,农杆菌菌液(OD600值为0.4)侵染20 min,共培养4 d,在含30 mg/L卡那霉素(Km)的培养基上诱导不定芽,生根培养基中Km的适宜浓度为10 mg/L。

关 键 词:河北杨  根癌农杆菌  GV3101  35S-DAS5  遗传转化体系

Establishment of System for Genetic Transformation of Agrobacterium tumefaciens Mediated Transformation of Populus hopeiensis with Growth-promoting Gene
Wang Peiya , Yang Hui , Yang Tao , Zhang Jun , Guo Qi , Qiang Weiya , Zhou Jianping.Establishment of System for Genetic Transformation of Agrobacterium tumefaciens Mediated Transformation of Populus hopeiensis with Growth-promoting Gene[J].Biotechnology Bulletin,2012,0(3):141-147.
Authors:Wang Peiya  Yang Hui  Yang Tao  Zhang Jun  Guo Qi  Qiang Weiya  Zhou Jianping
Institution:2,3(1Institute of Biology,Gansu Academy of Sciences,Lanzhou 730000;2 Industrial Microbiology Engineering Research Center in Gansu,Lanzhou 730000;3Gansu Academy of Sciences,Lanzhou 730000;4College of Life Science,Lanzhou University,Lanzhou 730000)
Abstract:Several factors affecting transformation of Populus hopeiensis were studied by infecting with Agrobacterium tumefaciens GV3101,such as pre-culture time,adding AS,Agrobacterium concentration,infection time,co-culture time,concentration of kanamycin used for selection and adding time,of which the frequency of kanamycin resistance buds was used as measure standard.The superior transformation system was as following,pre-cultured 2-4 days and then infected 20 minutes in Agrobacterium diluted(OD600=0.4),co-cultured 4 days,transgenic tissues and shoots were selected with 30 mg/L Km and rooting stage with 10 mg/L Km.
Keywords:Populus hopeiensis Agrobacterium tumefaciens GV3101 35S-DAS5 Genetic transformation
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