中国兽医科学2017，Vol.47Issue(7)：897-902,6.DOI:10.16656/j.issn.1673-4696.2017.07.016
猪IL-4稳定表达细胞系的建立
Establishment of a cell line stably expressing porcine IL-4
摘要
Abstract
This study was conducted to establish cells lines stably expressing porcine IL-4.The cloned porcine IL-4 gene consists of 402 bp,and the ORF of porcine IL-4 encodes a protein of 133 amino acids.The IL-4 polypeptide had an estimated molecular mass of 15.02 ku,PI of 8.64 and displayed a high homology to that of Bostaurus(80.5％),Ovisaries(76.7％),Equuscaballus(72.7％),Feliscatus (72.9％),Canis lupus familiaris(72.0％),Homo sapiens(69.2％) and Mus musculus(41.5％),respective1y.After the digestion of EGFP-tagged pIRES2-EGFP vector by EcoR Ⅰ and BamH Ⅰ enzymes,the ORF of porcine IL-4 was inserted and named as pIRES2-EGFP-pIL-4.Then the pIRES2-EGFP-pIL-4 was transfected into porcine intestinal epithelial ceil line(IPEC-J2),which was then screened by G418 to obtain the cell line stably expressing porcine IL-4.Consequently,the expression of IL-4 mRNA in transfected cells was significantly increased compared to control cells at hour 24 after transfection(P＜0.01) and transfected cells also showed high expression of green fluorescent.After G418 selection and expansion(positive clone) for 30 passages,the expression of IL-4 mRNA was increased by 200 times to controlcompared cells.We concluded that the eukaryotic expression vector pIRES2-EGFP-pIL-4 was constructed successfully and a cell lines that show the stable expression of porcine IL-4 was established,which laid the foundation for further studies of its functions.关键词
猪/IL-4/真核表达/稳定表达细胞系Key words
pig/IL-4/eukaryotic expression/stable expression of cell lines分类
农业科技引用本文复制引用
李俊敏,蔡彬,雍艳红,巩栋梁,冯园,何玉林,巨向红..猪IL-4稳定表达细胞系的建立[J].中国兽医科学,2017,47(7):897-902,6.基金项目
国家自然科学基金项目(31101862 （）
31472243) （）
