绵羊白细胞内的慢病毒cDNA GAG基因PCR检测  

PCR Detection of Lentivirus cDNA GAG Gene in the White cells of sheep

作　　者：张辉[1] 阿合买提.买买提 简子健[1] 王远志[2] 

机构地区：[1]新疆农业大学动医学院,乌鲁木齐830052 [2]石河子大学分子生物学研究室,石河子832003

出　　处：《中国农学通报》2005年第4期7-9,38,共4页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金(30160065)

摘　　要：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基因全长,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,所测序列与Genbank中的基因序列进行比对,所克隆序列与慢病毒gag基因序列一致。实验表明:在绵羊的白细胞慢病毒的cDNAgag基因的PCR检测是可行的。Taking on genome of sheep periblood white cell as templete, we amplified Ovine lenti virus gag gene, and then cloned this fragment into pBS-T vector. The ligased product was transformed to E.coli DH5α. The positive recombinant was screened on Amp/IPTG/X-Gel plates and identified by restriction endonuclease XhoI, clony PCR and sequencing. Analised object gene sequences, the sequenced fragment is consistent with ovine lentivirus gag gene . we drew a conclusion that PCR Detection of Lentivirus DNA gag Gene in the White cells of sheep is feasible.

关 键 词：PCR检测 CDNA GAG基因 细胞内 GENBANK GAG基因 基因序列 PCR技术 绵羊慢病毒 感受态细胞 基因克隆 IPTG 初步鉴定 白细胞 基因组 外周血 重组子 gel 全长 酶切 行比 

分 类 号：S858.28[农业科学—临床兽医学] Q78[农业科学—兽医学]