摘要
以猪肾细胞(PK-15)为材料,比较真核细胞膜蛋白提取试剂盒、超声破碎法、反复冻融法、低渗裂解法4种不同方法对PK-15细胞膜蛋白提取的影响,结果表明,通过超声破碎的膜制备方法提取的PK-15细胞膜蛋白较理想。用此方法提取PK-15细胞膜蛋白,经SDS-PAGE后转印NC膜,利用病毒铺覆蛋白印迹技术(VOPBA)鉴定CSFV病毒的细胞受体。结果发现,PK-15细胞膜上有3种能与CSFV结合的蛋白质,分子量在90～100 ku之间,暗示这些蛋白可能是CSFV受体或者受体复合物。
The aim of this study was to compare the effects of four methods, Mem-PER Eukaryotic Membrane Protein Extraction Reagent Kit, freezing-thaw, ultrasonic treatment and low permeability cracking method, on extracting PK-15 cell membrane proteins. The results showed that ultrasonic treatment was more ideal for the extraction of membrane proteins in PK-15 cell. After SDS-polyacrylamide gel electro- phoresis （SDS-PAGE） , the PK-15 cell membrane proteins were transferred to nitrocellulose membrane, and virus overlay protein binding assay （VOPBA） was performed to identify the receptor protein for classical swine fever virus （CSFV）. There were three cellular proteins of 90 ku to 100 ku that could bind CSFV in PK-15 cells, indicating that these proteins might be putative receptors or a part of receptor complex for CSFV.
出处
《畜牧与兽医》
北大核心
2011年第5期1-4,共4页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31001062
30571376)
南京农业大学青年科技创新基金(KJ08016)
关键词
PK-15细胞
膜蛋白提取
猪瘟
病毒受体
PK-15 cell
membrane protein extraction
classical swine fever virus （CSFV）
virus receptor
