| PK-15细胞膜蛋白提取方法的比较及猪瘟病毒膜表面受体的初步鉴定 |
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| 引用本文： | 周斌,刘晓东,王凤鹃,刘珂,陈溥言.PK-15细胞膜蛋白提取方法的比较及猪瘟病毒膜表面受体的初步鉴定[J].畜牧与兽医,2011,43(5). |
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| 作者姓名： | 周斌 刘晓东 王凤鹃 刘珂 陈溥言 |
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| 作者单位： | 南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京,210095 |
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| 基金项目： | 国家自然科学基金，南京农业大学青年科技创新基金 |
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| 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。
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| 关 键 词： | PK-15细胞 膜蛋白提取 猪瘟 病毒受体 |
Comparison on different extraction protocols of PK-15 cell membrane proteins and identification of cellular receptors for classical swine fever virus |
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| ZHOU Bin,LIU Xiao-dong,WANG Feng-juan,LIU Ke,CHEN Pu-yan.Comparison on different extraction protocols of PK-15 cell membrane proteins and identification of cellular receptors for classical swine fever virus[J].Animal Husbandry & Veterinary Medicine,2011,43(5). |
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| Authors: | ZHOU Bin LIU Xiao-dong WANG Feng-juan LIU Ke CHEN Pu-yan |
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| Institution: | ZHOU Bin,LIU Xiao-dong,WANG Feng-juan,LIU Ke,CHEN Pu-yan (Key Laboratory of Animal Diseases Diagnosis and Immunology,Ministry of Agriculture,Nanjing Agricultural University,Nanjing 210095,China) |
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| Abstract: | 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 electrophoresis(SDS-PAGE),the PK-15 cell membrane proteins were transferred to nitrocellulose membrane,and virus overlay protein b... |
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| Keywords: | PK-15 cell membrane protein extraction classical swine fever virus(CSFV) virus receptor |
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