中国兽医科学2017，Vol.47Issue(4)：448-454,7.DOI:10.16656/j.issn.1673-4696.2017.04.007
犬干扰素α4在CHO细胞中的稳定表达
Stable expression of canine interferon alpha4 in CHO cells
摘要
Abstract
In order to obtain canine interferon alpha4(CaIFN-α4) with high activity,we prepared the fusion gene of canine interferon alpha4 and green fluorescent protein(GFP),CaIFN-alpha4-GFP,containing the canine interferon alpha4 and GFP with two GGGGS-five peptide (2G4S).The recombinant expression plasmidpL-CaIFN-α4-GFPwas constructed bycloningthefusiongene(CaIFN-α4-GFP)into pL-eGFPeukaryotic expression vector.Recombinant lentivirus,vCaIFN-α4-GFP,was harvested following transfection of 293FT cells with pL-CaIFN-α4-GFP eukaryotic expression plasmid and other three lentivirus plasmids.Finally,we successfully established Chinese hamster ovary(CHO) cell expressing CaIFN-α4-GFP through infecting the CHO cells with the harvested recombinant lentivirus,followed by screened with blast fungus element,flow cytometry sorting and limited dilution transfection cell cloning.Green fluorescence could be observed in the established CHO cells,demonstrating the expression of interferon.The CHO cell culture supernatant had the antiviral activity agianst vesicular stomatitis virus that was detected with MDCK and MDBK,and the antiviral activities were 1.39× 105U/mL and 2.19 × 104 U/mL,respectively.This study successfully developed CHO cells expressing the canine interferon alpha4 with activity,which laid the experimental foundations for the development of the production system of dogs interferon alpha4 with high activity.关键词
犬干扰素α4(CaIFN-α4)/CHO细胞/真核表达/慢病毒/绿色荧光蛋白Key words
canine interferon α4 (CaIFN-α4)/CHO cell/eukaryotic expression/lentivirus/green fluorescent protein(GFP)分类
医药卫生引用本文复制引用
王晶宇,安欣,楼晶瑶,姚火春,欧阳伟,王永山,夏兴霞,诸玉梅,王晓丽,潘群兴,孟凯,毕振威..犬干扰素α4在CHO细胞中的稳定表达[J].中国兽医科学,2017,47(4):448-454,7.基金项目
国家重点研发计划项目(2016YFD0501000) （2016YFD0501000）
公益性行业(农业)科研专项(201303042) （农业）
江苏省农业科技自主创新资金项目[CX(15)1065] （15）
