摘要
为真核表达鹅细小病毒(GPV)融合蛋白VP3基因,本研究通过PCR扩增GPV FY89株VP3基因,并将其克隆至杆状病毒转移载体pFastBac/HBM-TOPO中,经双酶切、PCR及测序鉴定正确后,转化感受态细胞DH10Bac,提取重组杆状病毒穿梭质粒rBacmid-VP3,将经PCR鉴定正确的rBacmid-VP3转染Sf9昆虫细胞,连续传代后,细胞病变明显。SDS-PAGE分析结果表明成功表达了分子质量约为61ku的VP3蛋白;Western blotting分析结果表明重组VP3蛋白具有良好的抗原性。
The fusion VP3 gene sequence of goose parvovirus （GPV） was amplified by PCR using a pair of specific primers from GPV FY89 strain. The PCR product was cloned into pFastBac/HBM TOPO vector and transformed into DH10Bac cells. Recombinant Bacmid-VP3 was transfected into Sf9 insect cells,and a fusion protein about 61 ku was expressed and identified by SDS-PAGE analysis. The antigenicity of the recombinant VP3 protein was confirmed by Western blotting.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第9期52-55,共4页
China Animal Husbandry & Veterinary Medicine
基金
黑龙江省财政厅省属科研院所自拟专项基金
