人白细胞介素-37慢病毒载体的构建与包装  

Construction and packaging of human interleukin-37 lentiviral vector

作　　者：宋改环[1] 高雪明[2] 蒋雯雯[2] 李燕[1,2] 

机构地区：[1]宁波大学医学院附属医院,宁波315020 [2]宁波大学医学院

出　　处：《现代实用医学》2017年第5期568-570,556,共4页Modern Practical Medicine

基　　金：浙江省自然科学基金(LY17H010001);宁波市自然科学基金(2016A610088);宁波市自然科学基金(2015A610233);宁波市科技惠民项目(2015C50018)

摘　　要：目的探讨人白细胞介素(IL)-37慢病毒表达载体构建和包装。方法以pcDNA3.1(+)-IL-37为模板,用特异性引物聚合酶链式反应(PCR)扩增出IL-37全长编码序列;与载体pLVX-IRES-Green1酶切并连接,大量扩增纯化重组质粒及包装质粒;以适当比例共同转染293T细胞生产慢病毒;收集并浓缩慢病毒,检测慢病毒滴度及IL-37的表达。结果所得病毒的滴度达到5×10^(12) TU/ml;病毒感染A549细胞后,可以在细胞内稳定表达出IL-37 mRNA。结论本实验成功构建出IL-37慢病毒表达载体,并包装获得了高滴度的病毒上清。所包装病毒上清具有感染力,在真核细胞中可以表达IL-37。Objective To investigate the construction and packaging of human interleukin （EL ） -37 lentiviral vector.Methods The full-length coding sequence of IL-37 was amplified by PCR with pcDNA3.1 （＋）-EL -37 as a template.The sequence was cloned into pLVX- IRES-Greenl vector. The recombinant plasmids and packaging plasmids were am-plified, and the lentiviral was made by transfecting 293T cells in appropriate proportion. Lentivirus was collected and concentrated. Lentiviral titer and JL-31 expression were detected. Results After determination of titer and IL-37 ex-pression, it was found that the titer of the obtained virus reached 5 ×10^12 TU / ml. After infection with A549 cells, EL -37 mRNA could be stably expressed in the cells. Conclusions The IL-37 lentiviral expression vector was successfiilly constructed and the virus supernatant was obtained by packaging. The packaged virus supernatant is infectious and can express BL -37 in eukaryotic cells.

关 键 词：白细胞介素-37 慢病毒载体 构建 包装 

分 类 号：Q782[生物学—分子生物学] R446[医药卫生—诊断学]