| 重组犬IFN-γ在HEK293T细胞中的表达 |
| |
| 引用本文： | 靳慧君,李楠,郭常春,韩冬梅,王微,仲飞.重组犬IFN-γ在HEK293T细胞中的表达[J].河北农业大学学报,2009,32(2). |
| |
| 作者姓名： | 靳慧君 李楠 郭常春 韩冬梅 王微 仲飞 |
| |
| 作者单位： | 1. 河北农业大学,动物科技学院,河北,保定,071001
2. 广州天王动物保健品有限公司,广东,广州,510800 |
| |
| 摘 要： | 为了建立一套犬IFN-γ(cIFN-γ)在HEK293T细胞中表达的方法,用伴刀豆球蛋白A(ConA)刺激犬脾细胞,通过RT-PCR方法从脾细胞中扩增犬IFN-γ基因,并将其克隆到pcDNA3.1A载体中,构建的真核表达载体pcDNA3.1A-cIFN-γ经磷酸钙介导转染HEK293T细胞进行表达。结果表明:克隆的犬IFN-γcDNA基因与GenBank上发表的序列一致,同源性100%。表达产物经Western-blot方法检测,证明克隆的犬IFN-γ基因能够在HEK293T细胞中进行表达,并且表达产物能够分泌到细胞外。
|
| 关 键 词： | 犬γ-干扰素 HEK293T细胞 表达 |
Expression of recombinant canine IFN-γ gene in HEK 293T cells |
| |
| JIN Hui-jun,LI Nan,GUO Chang-chun,HAN Dong-mei,WANG Wei,ZHONG Fei.Expression of recombinant canine IFN-γ gene in HEK 293T cells[J].Journal of Agricultural University of Hebei,2009,32(2). |
| |
| Authors: | JIN Hui-jun LI Nan GUO Chang-chun HAN Dong-mei WANG Wei ZHONG Fei |
| |
| Institution: | 1.College of Animal Science and Technology;Agricultural University of Hebei;Baoding 071001;China;2.Guangzhou Tianwang Animal Health Products Co.L td;Guangzhou 510800;China |
| |
| Abstract: | To establish the method of canine IFN-γ(cIFN-γ) expression in HEK293T cells,the canine spleen cells were stimulated by concanavalin A.The cDNA encoding cIFN-γ was amplified by RT-PCR from the stimulated cells.Then the cIFNγ gene was inserted into eukaryotic expression vector pcDNA3.1A.The recombinant pcDNA3.1A-cIFN-γ plasmids were transfected into HEK293T cells mediated by calcium phosphate.Results showed that the sequence of cIFN-γ cDNA amplified was identical to that published in GenBank,and the homology is 100%.The expressed products were detected by Western-blot,which indicated that cIFN-γ gene could be expressed in HEK293T cells and secreted from the cells. |
| |
| Keywords: | canine IFN-γ HEK293T cells expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
