重组5型腺病毒高效扩增的研究  

Efficient amplification of recombinant type-5 adenovirus vector

作　　者：曹洁玮[1] 李小鹰[1] 

机构地区：[1]解放军总医院南楼心血管一科,北京100853

出　　处：《中华老年心脑血管病杂志》2010年第3期260-262,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(30770900)

摘　　要：目的探讨高效扩增腺病毒载体的方法。方法采用人胚肾293细胞株扩增合有SERCA2a基因的重组5型腺病素(Ad-SERCA2a),采用半组织定量法测定病毒滴度,PCR和Western blot法检测SERCA2a基因表达。结果扩增纯化后腺病毒滴度为1.0×10^(12) PFU/ml,扩增病毒SERCA2a基因表达。结论采用人胚肾293细胞扩增重组腺病毒,经纯化后可获得高滴度病毒。Objective To investigate the method of efficient amplification of recombinant adenovirus vectors. Methods HEK 293 cells were used to amplify recombinant type-5 adenovirus vector containing SERCA2a gene（Ad-SERCA2a）. Infectious adenoviral titer was measured by the medi- an tissue culture infectious dose（TCID50）, SERCA2a expression was detected by PCR and Western blot. Results The titer of adenovirus after amplification and purification was about 1.0 × 10^12 PFU/ml,SERCA2a was expressed. Conclusions High titer of recombinant adenovirus vector can be obtained after amplification with HEK 293 cells and purification.

关 键 词：腺病毒科 聚合酶链反应 基因扩增 基因表迭 

分 类 号：R373[医药卫生—病原生物学]