| 摘要: |
| 为了构建犬瘟热病毒F基因真核表达载体,利用RT-PCR方法扩增出水貂源犬瘟热病毒分离株的F基因,将其克隆至pMD-18T载体上,用BamHⅠ和KpnⅠ进行双酶切,回收目的基因。将目的基因亚克隆至pcD-NA3.1（＋）中,获得真核重组质粒pcDNA3.1-CDVF。通过脂质体介导法将pcDNA3.1-CDVF转染至BHK-21细胞中,并利用间接免疫荧光试验和RT-PCR法检测pcDNA3.1-CDVF在BHK-21细胞中的表达情况。结果表明,真核重组表达质粒pcDNA3.1-CDVF构建成功,F基因可在BHK-21细胞中表达。 |
| 关键词: 水貂 犬瘟热病毒 F基因 真核表达 |
| DOI： |
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| Construction and expression of eukaryotic expression plasmid for F gene of canine distemper virus Isolated from mink |
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| Abstract: |
| In order to construct eukaryotic expression plasmid for F gene of canine distemper virus，the F gene of CDV strain isolated from mink was amplified by reverse transcription polymerase chain reaction (RT-PCR)．The amplified fragments were cloned into pMD-18T simple vector．Objective gene was obtained by digesting the cloning plasmid with BamHⅠand KpnⅠ,and the eukaryon recombinant expression plasmids was constructed by re-cloning the objective gene into pcDNA3.1(+) vector.Eukaryon recombinant expression plasmid pcDNA3.1-CDVF was transfected into BHK 21 cell by using lipoplast mediate method and it was proved with indirect fluorescent antibody technique and RT-PCR detection that the objective gene was successfully expressed in BHK-21 cell.The results of the study provided fundamental information for the further study on the prevention of mink CD and new generation vaccine. |
| Key words: mink canine distemper virus F gene eukaryotic expression |
