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| 猪传染性胃肠炎病毒S蛋白主要抗原位点在昆虫细胞中的表达 | |
| 引用本文： | 李菁,何孔旺,杨倩.猪传染性胃肠炎病毒S蛋白主要抗原位点在昆虫细胞中的表达[J].南京农业大学学报,2009,32(1). |
| 作者姓名： | 李菁 何孔旺 杨倩 |
| 作者单位： | 1. 南京农业大学动物医学院,江苏南京210095;江苏省农业科学院兽医研究所,江苏南京210014 2. 江苏省农业科学院兽医研究所,江苏南京,210014 3. 南京农业大学动物医学院,江苏南京,210095 |
| 摘 要： | 用RT-PCR方法扩增届猪传染性胃肠炎病毒S基因主要抗原位点A、D,将其亚克隆至供体质粒pFastBacl,得到重组转移质粒pFastBael-TS1,转化到Escherichia coli DH5α中,提取阳性克隆质粒再转化到含穿梭载体bacmid的E.coliDHl0Bac中,发生转座作用,经蓝白斑筛选得到重组穿梭载体Bacmid-TS1,经PCR鉴定后再转染Sf9昆虫细胞,经空斑筛选得到纯化的重组病毒rAcV-Bac-FS1,经1FA、SDS-PAGE和Western-blot鉴定,重组杆状病毒TS1蛋白(相对分子质量约43 000) 在Sf9昆虫细胞中得到表达. |
| 关 键 词： | 猪传染性胃肠炎病毒 S蛋白 重组杆状病毒 表达 |
Expression of the main sites of transmissible gastroenteritis virus spike protein in insect cells |
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| LI Jing,HE Kong-wang,YANG Qian.Expression of the main sites of transmissible gastroenteritis virus spike protein in insect cells[J].Journal of Nanjing Agricultural University,2009,32(1). | |
| Authors: | LI Jing HE Kong-wang YANG Qian |
| Institution: | 1.College of Veterinary Medicine;Nanjing Agricultural University;Nanjing 210095;China;2.Institute of Veterinary Medicine;Jiangsu Academy of Agricultural Sciences;Nanjing 210014;China |
| Abstract: | The main sites of transmissible gastroenteritis virus spike gene A and D was amplified by RT-PCR and was cloned into the contribution vector pFastBac1 to get the recombinant plasmid pFastBac1-TS1.pFastBac1-TS1 was then introduced into Escherichia coli DH10Bac,which included a shuttle vector,bacmid.By site-specific transposition,S gene sites A and D were integrated into bacmid,and a recombinant shuttle vector was constructed,named Bacmid-TS1.The recombinant bacmid,which was identified by PCR,was transfected ... |
| Keywords: | transmissible gastroenteritis virus spike protein recombinant baculovirus expression |
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