绵羊白细胞内的慢病毒cDNA gag基因的PCR检测

对5只新疆卡拉库尔羊进行绵羊慢病毒免疫琼脂凝胶扩散检测试验,3只阳性,2只阴性,然后采用PCR技术,以绵羊外周血白细胞基因组为模板,扩增绵羊慢病毒gag基因,将该基因克隆到pBS-T载体上,将重组子转入感受态细胞DH5α,用IPTG/X-gel筛选阳性菌落,通过菌落PCR,酶切初步鉴定后测序,结果表明PCR的检测结果和AGIDT的结果一致,所测序列与Genbank中的Visna病毒的gag基因序列有一定的同源性.

PCR Detection of Lentivirus DNA gag Gene in Ovine Leucocytes White Blood Cells

5 Xinjiang Kalakuer sheep were tested by AGIDT,among which three sheep were positive, other two negative. Ovine lentivirus gag gene from peripheral blood cells in the five sheep,in which the sheep were collected, were used as template and amplified by PCR. Then the amplified fragments were cloned into pBS-T vector. The legated product was transformed into E.coli DH5α. The positive recombinant was screened on Amp/IPTG/X-Gel plates and identified by restriction endonuclease XhoI, clony PCR and sequencing. The result shows that the findings of PCR are consistent with that of AGIDT and there is some homologue between the gag gene sequence of OPPV and Visna virus in the Genbank.