期刊论文

Expression and Antibody Preparation of Porcine Adenovirus Type 3 Protease Protein in Escherichia coli

中文

中国畜牧兽医

1671-7236

2019

46

6

1764-1773

10.16431/j.cnki.1671-7236.2019.06.024

201710546001:国家级大学生创新创业训练计划项目 17B041:湖南省教育厅优秀青年项目 ZXXQ201701:校企合作项目 18H03:校企合作项目 16D20:***专项

本校为第一机构

试验旨在研究猪腺病毒3型(PADV3) Protease蛋白在大肠杆菌中的表达,并制备该蛋白的多克隆抗体.利用PCR扩增PADV3 Protease基因,构建重组原核表达载体pET28a-PADV3-Protease和真核表达载体pEGFP-PADV3-Protease,采用双酶切和测序鉴定;将原核表达载体pET28a-PADV3-Protease转化大肠杆菌BL21(DE3)感受态细胞得到重组原核表达菌株,经IPTG诱导收集蛋白,采用SDS-PAGE和Western blotting鉴定,将目的蛋白纯化后与佐剂乳化制备免疫原免疫家兔,制备Protease蛋白多克隆抗体;将真核表达载体pEGFP-PADV3-Protease转染HEK293细胞经G418筛选建立稳定表达EGFP-Protease融合蛋白的细胞系,以该稳定表达细胞系为包被...

This study was aimed to investigate the expression of porcine adenovirus type 3(PADV3)Protease protein in Escherichia coli(E.coli),and prepare the polyclonal antibody of Protease protein.PADV3 Protease gene was amplified by PCR,the recombinant prokaryotic expression vector pET28 a-PADV3-Protease and eukaryotic expression vector pEGFP-PADV3-Protease were constructed and identified by double enzyme digestion and sequencing.The recombinant prokaryotic expression vector pET28 a-PADV3-Protease was transformed into E.coli BL21(DE3),the positive recombinant prokaryotic expression strain was induced b...