人和鼠源性RANTES基因的克隆和腺病毒表达载体的建立  

Cloning the human and murine origin RANTES genes and Constructing the Adenovirus Vectors

作　　者：史业辉[1] 曹水[1] 刘虹[1] 李慧[1] 任秀宝[1] 

机构地区：[1]天津医科大学肿瘤医院,天津300060

出　　处：《天津医科大学学报》2004年第1期1-4,共4页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助 (编号 :30300438)

摘　　要：目的 :克隆人和小鼠的活化T细胞表达与分泌调节基因 (RANTES基因 )并分别构建腺病毒表达载体。方法 :利用RT -PCR方法从人的扁桃体组织和小鼠的脾细胞中克隆RANTES基因并将其克隆入T -easy保存载体中 ,使用Stratagene公司的ADEasy -XL腺病毒载体组装试剂盒建立表达该基因的腺病毒表达载体并使用CsCl2 密度梯度离心法浓缩病毒悬液。空斑试验显示病毒的滴度为2×1011cfu/ml。结果 :该实验成功地克隆了人和小鼠的RANTES基因并分别建立了表达该基因的腺病毒表达载体。结论Objective:To clone the human and murine origin RANTES genes and construct the adenoviral expression vectors.Methods:Using RT-PCR to clone the cDNA of human and murine origin RANTES genes and transduct into T-easy vector.Then constructed the adenˉoviral vectors were expressing the RANTES genes using the ADEasy-XL Adenoviral Vector System of Stratagene.After processed CsCl 2 density gradient centrifugation and plaque assays,2×10 11 cfu/ml virus are obtained.Results:The trial cloned the human and murine origin RANTES genes and constructed the adenoviral expressing vectors with high titer was successˉful.Conclusion:The construction of the adenoviral vectors expressing the human and murine origin RANTES genes with high titer make the basis of further functional trials.

关 键 词：活化T细胞表达与分泌调节基因(RANTES基因) 病毒载体 空斑试验 

分 类 号：R392.11[医药卫生—免疫学]