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Position: Home > Articles > Optimizing Transient Genetic Transformation Method on Arabidopsis Plants Mediated by Agrobacterium tumefaciens Journal of Northeast Forestry University 2016,44 (6) 41-44+83

一种基于农杆菌介导的拟南芥瞬时转化技术优化

作  者:
郭勇;王玉成;王智博
单  位:
林木遗传育种国家重点实验室(东北林业大学)
关键词:
拟南芥;GUS基因;瞬时表达;实时定量;农杆菌介导转化
摘  要:
利用根癌农杆菌(Agrobacterium tumefaciens)介导的拟南芥瞬时转化体系影响因素来确定最佳转化条件。以生长15日龄的拟南芥幼苗为试验材料,以转化p CAMBIA1301空载体的根癌农杆菌EHA105为目的菌株进行瞬时转化。研究了吐温20、菌液OD值、乙酰丁香酮(AS)及转化时间等对拟南芥瞬时转化效率的影响。结果表明:以体积分数为0.05%的吐温、菌液OD_(600)值为1.0和120μmol/L的AS侵染拟南芥2.5 h后,再共培养72 h,能够得到高瞬时转化效果。
译  名:
Optimizing Transient Genetic Transformation Method on Arabidopsis Plants Mediated by Agrobacterium tumefaciens
作  者:
Guo Yong;Wang Yucheng;Wang Zhibo;State Key Laboratory of Forest Genetics and Tree Breeding,Northeast Forestry University;
关键词:
Arabidopsis thaliana;;GUS gene;;Transient expression;;Real-time quantitative;;Agrobacterium mediated transformation
摘  要:
We studied the optimal transformation conditions for transient genetic transformation of Arabidopsis mediated by Agrobacterium tumefaciens. We transformed Agrobacterium EHA105 carrying an empty pC AMBIA1301 into the 15-day Arabidopsis plants for genetic transformation research. We studied the effects on transient expression including the concentration of the Agrobacterium,Tween-20,and acetosyringone( AS),and transformed time. Under the conditions of 1.0 OD of Agrobacterium,0.05% of Tween-20,120 μmol / L of AS,transformed for 2.5 hours,and co-cultured for 3 days,the transient transformation efficiency is highest.

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