摘要
采用Gateway技术将鸡传染性法氏囊病病毒(IBDV)Gx株和Gt株vp2基因分别克隆到载体pDEST-32构建诱饵载体,通过酶切和PCR鉴定并测序正确后,用醋酸锂法转化酵母菌株MaV203,通过缺陷型平板筛选和X-gal颜色筛选检测诱饵载体有无自激活作用。结果表明,成功构建了诱饵载体pDEST-32-Gxvp2和pDEST-32-Gtvp2,并证明其在酵母双杂交系统中无自激活作用。为下一步利用酵母双杂交方法从鸡法氏囊B淋巴细胞和鸡胚成纤维细胞cDNA表达文库中筛选与强、弱毒VP2诱饵载体作用的分子奠定了基础。
The fragments of infectious bursal disease virus （IBDV） GxVP2 and GtVP2 were cloned into pDEST-32 vector using Gateway technology and confirmed by restriction enzyme digestion, PCR and sequence analysis. Bait vectors were trans-formed into the yeast strain MaV203 by PEG/LiAC method and their self-activations were tested by both selective plate and X-gal color assay. The results showed that the bait vectors pDEST-32-GxVP2 and pDEST-32-GtVP2 had no autonomous activation, therefore could serve as baits in the screening of chicken bursal B homeocyte expression library and chicken embryo tibroblasts expression library to trap the interaction proteins using yeast two-hybrid system.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2008年第1期1-4,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家973项目(2005CB523202)
关键词
IBDV
酵母双杂交
诱饵载体
自激活
IBDV
yeast two-hybrid system
bait vector
autonomous reporter activation
