农杆菌介导的pCB302-3载体在本氏烟中瞬时表达条件优化

Optimizing agroinfiltration-mediated transient expression in Nicotiana benthamiana using pCB302-3 vector

以GFP为报告基因构建植物表达载体pCB302-3-GFP,采用农杆菌叶片注射法对其在本氏烟叶片中瞬时表达条件进行优化。分析基因沉默抑制子p19、菌液不同浓度以及侵染时间对GFP荧光强度的影响,结果显示,当农杆菌菌液D600为0.8-1.0并与含有p19基因的载体共注射条件下,农杆菌注射3-5d后本氏烟草叶片表现出很强的绿色荧光,实现了GFP在本氏烟草中的高效瞬时表达。

pCB302-3 is a plant mini-binary stable transSormation vector. To investigate the transient expression of pCB302-3 vector in plant,GFP was inserted into pCB302-3 vector as a reporter gene,and various 5actors including density of Agrobacteriurn cell, supplementation of gene silencing suppression p19 and days post infiltration were optimized based on agroinfiltration method in Nicotiana bentharniana leaves. Results showed that high levels of GFP expression were observed in N. bentharniana leaves 3-5 d aster infiltration by Agrobacterium cell suspension contained pCB302-3-GFP with an optical density （D600） of 0.8-1.0 co-infiltrated with p19 gene.