摘要
为建立从患有非典型新城疫的蛋鸡组织细胞中提取、纯化热休克蛋白HSP70抗原肽复合物的方法,采取三步蛋白纯化法从非典型新城疫蛋鸡肝脏组织中提取、纯化HSP70抗原肽复合物。经分级硫酸铵盐析后,依次用ConA-Sepharose亲和层析和DEAE-Sephacel离子交换层析分离纯化。所得蛋白经SDS-聚丙烯酰胺凝胶电泳和Western blotting进行蛋白分子量及性质鉴定,Bradford法测定蛋白浓度。分离、纯化得到的蛋白经SDS-PAGE、银染鉴定为单一带,分子量为70ku;Western blotting结果证实为HSP70。每克新鲜肝脏细胞最终获得90～110μgHSP70。使用本分离纯化方法可获得高纯度HSP70抗原肽复合物。
To develop the method to purify HSP70 antigen peptide complex from tissues of layers infected with atypical Newcastle disease. The mixed protein was precipitated by ammonium sulfate,and separated by affinity chromatography on Con A-Sepharose and DEAE-Sephacel exchange chromatography. The molecular weight and identity of the HSP70 glyeoprotein was confirmed by SDS-PAGE and Western blotting,the concentration was measured by the means of Bradford. A sharp stained protein band with a molecular weight of about 70 ku was obtained and confirmed to be HSP70 by Western blotting,90 to 110 μg of HSP70 could be obtained from 1 g of chicken tissues. The pure HSP70-peptide complexes could be obtained by this method.
出处
《中国家禽》
北大核心
2009年第16期19-21,共3页
China Poultry
