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高致病性猪繁殖与呼吸综合征病毒TA-12株ORF6基因的克隆、鉴定及真核表达

, PP. 159-163

Keywords: 高致病性猪繁殖与呼吸综合征病毒,ORF6蛋白,真核表达

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Abstract:

旨在获得HP-PRRSVTA-12(GenBank登录号:HQ416720)分离株ORF6蛋白的高效表达,将HP-PRRSVTA-12分离株的ORF6基因克隆到载体PfastBacHTB,并经转座进入E.coli含有杆状病毒质粒Bacmid的DH10BacTM细胞株,通过蓝白斑筛选和PCR鉴定出阳性重组杆粒,再将提取的重组杆粒Bacmid-ORF6转染昆虫细胞sf9,使其表达ORF6蛋白。经Westernblot鉴定,在感染重组杆状病毒后第4天ORF6蛋白表达量最大,达到0.05μg/μL。表达的ORF6蛋白主要存在于细胞质中。

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