CXCR4-shRNA重组腺病毒表达载体的构建及鉴定  

Construction and identification of recombinant adenovirus expression vector of short hairpin RNA for CXCR4

作　　者：陆航[1,2] 孙巨峰[2] 刘学政[1] 

机构地区：[1]广西医科大学人体解剖学教研室,南宁530021 [2]辽宁医学院附属第一医院

出　　处：《山东医药》2011年第46期22-23,共2页Shandong Medical Journal

基　　金：辽宁省自然科学基金资助项目(201102124)

摘　　要：目的选择性构建CXC趋化因子受体4(CXCR4)的特异RNA干扰载体。方法在GenBank中获取CXCR4基因的核苷酸序列,设计并合成3条短发夹RNA(shRNA)的DNA序列,并克隆到PL-Dest腺病毒骨架载体(shpAd/PL-Dest)中,组成人趋化因子受体重组腺病毒基因沉默子(shpAd-hCXCR4-eGFP),采用酶切、PCR和DNA测序方法验证。将验证后3种沉默子经酶切后转染至HEK293A细胞包装病毒,得到CXCR4的腺病毒表达载体。结果构建的shRNA-Ad-hCXCR4-eGFP 3种基因沉默子序列正确并成功克隆到腺病毒表达载体上。结论成功构建CXCR4-shRNA腺病毒表达载体。Objective To selectively construct of specific adenovirus expression vector of RNA interierence ior CXCR4. Methods Genome sequences of CXCR4 gene were retrieved from GenBank, then three cDNAs were designed and synthesized coding expression of small hairpin RNAs （shRNA） for CXCR4 gene, and after renaturation, it was cloned into the framework plasmid （shpAd/PL-Dest） of adenovirus to construct the CXCR4-shRNA expression vector shpAd-h CXCR4-eGFP. The recombinant expression vectors were identified by enzyme cutting method, PCR and the sequence analysis. After verification, the three silencers which were cut by enzyme were transfected into HEK293A cells to obtain CX- CR4 adenoviral expression vector. Result The three synthetic sequences were completely correct and successfully cloned to adenoviral expression vector. Conclusion The adenoviral expression vector of CXCR4-shRNA has been constructed successfully.

关 键 词：CXC趋化因子受体4 RNA干扰 重组腺病毒载体 人胚肾293A细胞 

分 类 号：R656.6[医药卫生—外科学]