大熊猫犬冠状病毒部分纤突蛋白基因的扩增与序列分析

Amplification and Sequence Analysis of Partial Spike Protein Gene of Canine Coronavirus Isolated from the Liver of a Giant Panda

大熊猫是我国特有的濒危物种,近年来人们在研究其生态环境、生理特点、繁殖性能、人工繁育等方面取得了显著成绩[1].然而,大熊猫疾病,特别是传染性疾病对大熊猫的生存构成了严重的威胁.

Liver material of a dead giant panda and different generations of CCV DXMV strain isolated from it were used to amplify partial spike protein gene of canine coronavirus by nested PCR with primers of CCVF1,CCVR1 and CCVF2,CCVR2. 1 086 bp and 515 bp nucleotide sequences were acquired respectively. The fragments were purified, sequenced and analyzed with the software of DNASTAR and DNASIS. The results showed that the sequences amplified from the giant panda liver and 2,3,29 generations of CCV DXMV between primers of CCVF2 and CCVR2 were identical and suggested that the virus was successfully isolated from the giant panda liver. 1 047 bp nucleotides between primers of CCVF1 and CCVR1 were correctly sequenced from the 3rd generation of CCV DXMV and its homology with other CCV strains were 83.3%~100%, especially 100% with the CCV NVSL strain that isolated from USA.