ZHONG Liang. Construction,expression and identification of yeast two-hybrid bait vector pGBKT7-PML-V[J]. Journal of Chongqing Medical University, 2008,(3):257-260.
ZHONG Liang. Construction,expression and identification of yeast two-hybrid bait vector pGBKT7-PML-V[J]. Journal of Chongqing Medical University, 2008,(3):257-260. DOI: 10.13406/j.cnki.cyxb.2008.03.001.
Western Blotting分析证实酵母细胞表达诱饵蛋白。结论:成功构建了PML-V结合域的酵母诱饵表达载体
为进一步运用酵母双杂交技术筛选与之相互作用的蛋白奠定了基础。
Abstract
Objective:To construct the bait expression vector pGBKT7-PML-V of retinoic acid receptor variant protein
for screening the target proteins interacting with the bait protein through the yeast two-hybrid system.Methods:The fragments of PML-V binding domain was amplified by PCR
and then was cloned into the bait expression vector pGBKT7.After being verified by sequencing
the bait vector pGBKT7-PML-V was transformed into AH109 yeast cells.Then the expression of the bait protein was analyzed by Western Blotting.Toxicity
leakage and serf-activation of the bait protein were detected.Results: PML-V was amplified and cloned into pGBKT7 successfully.The bait vector was transformed into AH109 as well and no toxicity and self-activation were found.The expression of the bait protein was confirmed by Western Blotting.Conclusion:The bait expression vector of PML-V was constructed successfully
which layed the foundations for screening target proteins interacting with the bait protein using the yeast two-hybrid technique.