【摘要】：為提高表達(dá)豬圓環(huán)病毒2型(PCV2)ORF2基因的重組豬圓環(huán)病毒1型(PCV1)疫苗株(嵌合豬圓環(huán)病毒1-2,PCV1-2)在細(xì)胞內(nèi)的增殖水平,本研究采用PK-15細(xì)胞,應(yīng)用活細(xì)胞計(jì)數(shù)、間接免疫熒光及Taq Man熒光定量PCR等方法測(cè)定不同熱應(yīng)激溫度和持續(xù)時(shí)間處理后細(xì)胞生長(zhǎng)和病毒增殖的情況。結(jié)果顯示,PK-15細(xì)胞可耐受41℃和43℃的培養(yǎng)環(huán)境,并且與常規(guī)培養(yǎng)方法相比43℃條件下熱應(yīng)激處理12 h可顯著促進(jìn)PCV1-2在細(xì)胞內(nèi)的增殖,其病毒效價(jià)可達(dá)10~(6.0)TCID(_50)/m L以上。另外,熱應(yīng)激處理后病毒核酸拷貝數(shù)的動(dòng)態(tài)變化表明,熱應(yīng)激對(duì)PCV1-2復(fù)制的影響主要在接毒后24 h至48 h內(nèi)顯現(xiàn)。本研究確定了促進(jìn)PCV1-2增殖的熱應(yīng)激條件,為提高PCV1-2疫苗產(chǎn)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，

本文編號(hào)：2277728