中国兽医科学2025，Vol.55Issue(3)：389-395,7.DOI:10.16656/j.issn.1673-4696.2025.0026
猪CD40L的可溶性表达及其生物学活性检测
Soluble expression and biological activity detection of porcine CD40L
摘要
Abstract
In order to investigate the activation of soluble porcine CD40L(1 13-261 amino acids)on porcine peripheral blood mononuclear cells(PBMC),a plasmid containing porcine CD40L gene was constructed and expressed in Escherichia coli BL21(DE3).The purified recombinant CD40L protein was conjured with FITC and then hybridized with PBMC in situ.The biological activity of B cells stimulated by CD40L protein was studied by fluorescence microscopy and flow cytometry.SDS-PAGE and Western-blot results showed that the size of recombinant CD40L proteinwas about 14 kDa.The optimal expression conditions were 16 ℃,D600 nm value was 1.l,and the concentration of 0.5 mmol/L was induced by IPTG for 12 h.Flow cytometry analysis showed that the recombinant CD40L protein could bind to CD40 protein molecules on the surface of PBMC,and the recombinant CD40L protein could significantly activate B cells(P＜0.001).In conclusion,CD40L had the biological activity of promoting B cell activation,and CD40L can promote immune cell reaction,and have the immunomodulatory effect.This study provided a basis for the role of CD40L in important livestock species and animal models of human disease.关键词
猪CD40L/原核表达/生物学活性Key words
porcine CD40L/prokaryotic expression/biological activity分类
农业科技引用本文复制引用
贺艳艳,郭慧琛,孙世琪,穆素雨,敬晓棋..猪CD40L的可溶性表达及其生物学活性检测[J].中国兽医科学,2025,55(3):389-395,7.基金项目
国家成都农业科技中心地方财政专项(NASC2024KR06) （NASC2024KR06）
兰州市人才创业创新项目(2023-RC-3) （2023-RC-3）
