【作者】 李进； 王芹； 岳井银； 穆传杰； 唐卫生；

【Author】 WANG Qin, LI Jin, YUE Jingyin, MU Chuanjie, TANG Weisheng (Institute of Radiation Medicine,Chinese Academy of Medical Sciences and Peking Union Medical College,Tianjin 300192 China.)

【机构】 中国医学科学院中国协和医科大学放射医学研究所；

【摘要】 目的:构建携带人Rb94基因的重组腺病毒载体并扩增,为研究Rb94基因-放射治疗肿瘤奠定基础。方法:将已构建好的pENTRTM1A-Rb94穿梭质粒与腺病毒表达载体(pAd/CMV/V5-DESTTM)进行LR重组,构建含Rb94基因的腺病毒载体(Ad-Rb94)。Ad-Rb94在脂质体的作用下转染293A细胞进行扩增,制备重组腺病毒颗粒。用PCR方法对病毒鉴定和TCID50法测定其滴度。结果;构建的重组腺病毒载体含有Rb94基因,病毒的滴度为1.6×108pfu/ml。结论:成功构建携带人Rb94基因的重组腺病毒表达载体。更多还原

【Abstract】 Objective To construct the recombinant Rb94 gene adenovirus expression vector and explore a therapy approach for tumors using Rb94 gene with radiation. Methods The constructed shuttle vector pENTR1A-Rb94 and adenovirus vector pAd/CMV/V5-DEST was recombined by LR technique to construct the recombined Rb94 gene adenovirus expression vector(Ad-Rb94).The adenovirus stock was produced by cotransfection of 293A cells with Ad-Rb94 and lipofectamine .The identification of recombinant expression vector was performed by PCR method and the titer was measured by TCID50 .Results The recombinant adenovirus vector can express Rb94 gene. The adenovirus titer was 1.6×108pfu/ml. Conclusion The recombinant Rb94 gene adenovirus expression vector was constructed successfully.更多还原