热应激对嵌合猪圆环病毒1-2在PK-15细胞中增殖的影响
发布时间：2018-10-17 19:33
【摘要】：为提高表达猪圆环病毒2型(PCV2)ORF2基因的重组猪圆环病毒1型(PCV1)疫苗株(嵌合猪圆环病毒1-2,PCV1-2)在细胞内的增殖水平,本研究采用PK-15细胞,应用活细胞计数、间接免疫荧光及Taq Man荧光定量PCR等方法测定不同热应激温度和持续时间处理后细胞生长和病毒增殖的情况。结果显示,PK-15细胞可耐受41℃和43℃的培养环境,并且与常规培养方法相比43℃条件下热应激处理12 h可显著促进PCV1-2在细胞内的增殖,其病毒效价可达10~(6.0)TCID(_50)/m L以上。另外,热应激处理后病毒核酸拷贝数的动态变化表明,热应激对PCV1-2复制的影响主要在接毒后24 h至48 h内显现。本研究确定了促进PCV1-2增殖的热应激条件,为提高PCV1-2疫苗产量提供了方法。
[Abstract]:In order to improve the intracellular proliferation of recombinant porcine circovirus type 1 (PCV1) vaccine strain expressing porcine circovirus type 2 (PCV2) ORF2 gene (chimeric porcine circovirus 1-2pPCV1-2), PK-15 cells were used to count live cells. Indirect immunofluorescence and Taq Man fluorescence quantitative PCR were used to detect cell growth and virus proliferation after different heat stress temperature and duration. The results showed that PK-15 cells could tolerate 41 鈩，
本文编号：2277728
[Abstract]:In order to improve the intracellular proliferation of recombinant porcine circovirus type 1 (PCV1) vaccine strain expressing porcine circovirus type 2 (PCV2) ORF2 gene (chimeric porcine circovirus 1-2pPCV1-2), PK-15 cells were used to count live cells. Indirect immunofluorescence and Taq Man fluorescence quantitative PCR were used to detect cell growth and virus proliferation after different heat stress temperature and duration. The results showed that PK-15 cells could tolerate 41 鈩，
本文编号：2277728
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