人白介素24腺病毒载体的构建与鉴定  

The Construction and Identification of Recombinant Adenovirus Vector of hIL-24

作　　者：叶震敏[1] 张颂文[2] 缪竞诚[1] 盛伟华[1] 王伟录[1] 吴晓阳[2] 杨吉成[1] 

机构地区：[1]苏州大学医学院细胞与分子生物学教研室,江苏苏州215123 [2]昆山市第一人民医院普外科,江苏昆山215300

出　　处：《苏州大学学报（医学版）》2006年第1期44-47,共4页Suzhou University Journal of Medical Science

基　　金：苏州大学医学发展基金资助项目(EE134517);昆山市科技局社会发展科技基金资助项目(KS0405)

摘　　要：目的构建人白介素24(hIL_24)的腺病毒载体,获得hIL_24重组腺病毒子,为hIL_24进行肿瘤的基因治疗奠定基础。方法以pcDNA3.0_hIL_24重组质料为模板,PCR扩增hIL_24,酶切连接到带有GFP标记的pAdTrack_CMVR质粒上,PmeI线性化重组质粒pAdTrack_CMV_hIL_24,与腺病毒质粒pAdEasy_1共转化BJ5183细菌,获得重组腺病毒载体pAdEasy_1_pAdTrack_CMV_hIL_24,经Pacl线性化后转染QBI-293A包装细胞,收获腺病毒重组病毒子,RT_PCR和Western_blotting鉴定。结果测序结果显示hIL_24序列正确,RT_PCR和Western_blotting检测到了hIL_24的表达。结论成功构建人白介素24的重组腺病毒载体pAdEasy-1-pAdTrack_CMV_hIL_24,获得了人白介素24重组病毒子Ad_hIL_24。Objective The recombinant adenovirus vector of hlL-24 was constructed for carcinoma gene therapy. Methods The pAdTrack CMV-hIL-24 was constructed by PCR with pcDNA3.0-hIL- 24 recombinant plasmid as template, enzyme digestion and ligation. The pAdTrack-CMV-hIL-24 lineared by PmeI was co-transformed into BJ5183 with pAdEasy-1. The pAdEasy-1-pAdTrack-CMV- hIL-24 recombinant adenovirus vector was lineared with PacI and then transfected into the package cell OBI-293A. The hIL-24 recombinant adenovirus was obtained and identified by RT-PCR and Western-blotting. Results The hIL-24 was confirmed by sequencing, and the expression of hIL-24 was detected by RTPCR and Western-blotting. Conclusion The pAdEasy-1-pAdTrack-CMV-hIL-24 vector is constructed and the hIL-24 recombinant adenovirus（Ad-hIL-24）is obtained success fully.

关 键 词：人白介素24 腺病毒载体 构建 鉴定 

分 类 号：R392.2[医药卫生—免疫学]