中国兽医科学2025，Vol.55Issue(2)：197-202,6.DOI:10.16656/j.issn.1673-4696.2025.0015
CTAB对口蹄疫病毒样颗粒纯化及组装的影响
Effects of CTAB on purification and assembly of foot-and-mouth disease virus-like particles
摘要
Abstract
To determine the effect of cetyltrimethyl ammonium bromide(CTAB)on the clarification of E.coli bacterial sludge suspension and the assembly of foot-and-mouth disease virus-like particles(FMD VLPs).By adding CTAB into the E.coli bacterial sludge suspension of FMD virus structural protein,the changes of DNA and endotoxin contents were detected by nucleic acid electrophoresis and Limulus reagent color development method,respectively,to determine the clarifying effect of CTAB on the bacterial sludge suspension.Then the target protein was purified by His affinity chromatography gel method.The purified protein was assembled into FMD VLPs in a cell-free system and quantitatively analyzed by sucrose density gradient ultracentrifugation combined with high performance liquid phase analysis.After the addition of CTAB,the host DNA content in the supernatant was significantly reduced after homogenization(P＜0.01),and the concentration of harvested protein was not significantly changed after purification(P＞0.05),but the endotoxin content of harvested protein was significantly increased(P＜0.01),and the absolute Zeta potential of the supernatant and purified protein was significantly decreased(P＜0.05).The assembly amount of FMD VLPs was significantly decreased(P＜0.05).CTAB can precipitate DNA in a solution with a certain salt concentration,which is conducive to the removal of host DNA,but at the same time,it changes the charged property of the target protein,enhances the binding of the target protein with endotoxin,and reduces the stability of the assembly system,resulting in a significant decline in the assembly rate of FMD VLPs.关键词
CTAB/FMDV结构蛋白/DNA/内毒素/VLPsKey words
CTAB/FMDV structural protein/DNA/endotoxin/VLPs分类
农业科技引用本文复制引用
刘潇,余谦,田丽梅,杜平,蔺晓忠,严欢,李敏,李仙月,朱映新,杨雪,何清,钟丽君..CTAB对口蹄疫病毒样颗粒纯化及组装的影响[J].中国兽医科学,2025,55(2):197-202,6.基金项目
中央引导地方科技发展资金项目(202207AC110015) （202207AC110015）
云南省生物医药重大科技专项(202302AA310023) （202302AA310023）
云南省创新引导与科技型企业培育计划项目(202304AP080110) （202304AP080110）
云南省科技人才与平台计划项目(202405AC350099) （202405AC350099）
