中国畜牧兽医2017，Vol.44Issue(9)：2587-2592,6.DOI:10.16431/j.cnki.1671-7236.2017.09.007
猪白细胞介素18基因的克隆及真核表达重组质粒的构建
Cloning and Eukaryotic Expression Plasmid Construction of Porcine Interleukin-18 Gene
摘要
Abstract
To obtain recombinant eukaryotic expression plasmid of porcine interleukin-18(IL-18),the whole gene of porcine IL-18 gene was amplified from porcine spleen,lung and lymph nodes by RT-PCR and cloned into eukaryotic expression vector pZJ-1.The recombinant expression plasmid pZJ-IL-18 were identified by enzyme digestion and sequencing analysis,and was transfected into 293T cells.The expression of IL-18 was detected by Real-time PCR and Western lblotting in both gene and protein levels.The results showed that the eukaryotic expression plasmid of porcine IL-18 was constructed and could express transiently in 293T cells.Western blotting result confirmed that porcine IL-18 polyclonal antibody could react specifically with approximately 17 ku expression products,and indicated that IL-18 could express correctly and be responsive.This study constructed the eukaryotic expression plasmid of porcine IL-18 gene which could express transiently in 293T cells,and laid the foundation for studying function of IL-18.关键词
猪/IL-18基因/真核表达载体/克隆Key words
porcine/IL-18 gene/eukaryotic expression vector/clone分类
生物科学引用本文复制引用
石昂,时庆贺,李新果,王玉国,杨利,李双双,陈陆,王川庆..猪白细胞介素18基因的克隆及真核表达重组质粒的构建[J].中国畜牧兽医,2017,44(9):2587-2592,6.基金项目
国家自然科学基金(31272567、31772781) （31272567、31772781）
河南省高校科技创新人才支持计划(14HASTIT022) （14HASTIT022）
