重组腺相关病毒介导的nm23H_1载体的构建及鉴定  

Construction and Identification of Adeno-associated Virus Mediated Gene Transfer of nm23H_1

作　　者：李静[1] 陈刚[1] 高庆蕾[1] 邢辉[1] 邬素芳[1] 张阿丽[1] 卢运萍[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《华中科技大学学报（医学版）》2003年第4期421-424,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No 3984 0 0 0 9);国家杰出青年基金资助项目 (No 30 0 2 5 0 17)

摘　　要：目的　对载有肿瘤转移抑制基因nm2 3H1的重组腺相关病毒 (AAV)载体进行构建及鉴定。方法　用基因重组的方法构建含全长nm2 3H1的重组腺相关病毒骨架质粒 ,利用磷酸钙沉淀法将腺相关病毒载体系统共转染入病毒包装细胞 2 93细胞中 ,分别合成AAV nm2 3H1和AAV Laz,经PCR ,DNA测序 ,酶切鉴定 ,用斑点杂交法测定重组病毒的滴度。并用重组病毒感染卵巢癌细胞测定其转染效率。结果　成功构建并鉴定nm2 3H1,梯度均为 10 10 病毒分子 /ml,转染效率为 70 %。结论　构建AAV nm2Objective To construct and identify recombinant adeno associated nm23H1 virus. Methods Using DNA recombinant technique, main plasmid of AAV pUF1 nm23H 1 was constructed. Recombinant AAV nm23H 1/AAV Laz was produced by cotransfection of AAV system in package cell by phosphate calcium deposit method. The AAV nm23H 1 was identified by PCR, DNA sequencing and restriction digestion. The titer was measured by dot hybridization. The efficiency of transfection of AAV nm23H 1 into ovarian tumor cell SW626 was determined. Results AAV nm23H 1/AAV Laz were successfully constructed and identified. The titer were both 10 10 particles/ml. The efficiency of transfection was 70 %. Conclusion The development of AAV nm23H 1 provided a new tool for therapy of tumor metastasis.

关 键 词：NM23H1基因 腺相关病毒 肿瘤转移 斑点杂交法 

分 类 号：R73-37[医药卫生—肿瘤]