摘要
从疑似犬冠状病毒(CCV)症状病犬粪便中分离出一株病毒,合成通用引物(2Bp,4Bm)和CCV特异性引物(CCVF2,CCVR2)进行RT-PCR反应及限制性内切酶BstXⅠ酶切鉴定,并将纯化的PCR产物成功地克隆到pGEM-T-easy载体中。RT-PCR反应结果,BstXⅠ酶切鉴定结果以及重组质粒PCR鉴定结果均得到预计扩增片段。重组质粒测序结果与GenBank的CCV序列进行比对,与之相符,表明本次实验分离到的病毒为CCV。
A strain of virus was isolated from the fecal samples of a dog with clinical CCV illness. RTPCR Identification by use of 2 pairs of common primers 2 Bp and 4 Bm and specific primers CCVF2 and CCVR2 as well as BstX I degestion was carried out. PCR products were purified and cloned into the pGEM-T easy vector. The. result of RT-PCR, BstX Ⅰ digestion and recombinant plasmid amplified all gained the expect amplified segment. The result of sequencing shows that the PCR product is completely identical with that of the sequence searched in gene bank and indicated that the isolated virus is CCV.
出处
《西北农业学报》
CSCD
北大核心
2008年第2期33-37,42,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
公安部科技局资助项目(编号:20019829502)
