中国动物传染病学报2018，Vol.26Issue(1)：60-65,6.
环形泰勒虫TRAP-A蛋白酵母双杂交诱饵质粒的构建与鉴定
CONSTRUCTION AND IDENTIFICATION OF THE PGBKT7-TRAP-A PLASMID
摘要
Abstract
In the present study, the bait plasmid of pGBKT7-TRAP-A were constructed to screen the interactive proteins derived from the salivary glands of Hyalomma anatolicum anatolicum with Theileria annulata TRAP-A protein.The amplification primers were designed based on the sequence of TRAP-A adhensive domain of annulata merozoite.The fragment with the length of 576 bp was amplified and ligated into the pGBKT7vector. The pGBKT7-TRAP-A plasmid was confirmed using enzyme digestion and sequence analysis and then transformed into yeast cells Y2HGold. The TRAP-A fusion protein was expressed and examined for its autoactiviation and toxicity to yeast growth. Results indicated that the TRAP-A fusion protein was expressed in yeast cells Y2HGold without interruption of cellular growth. In addition, the recombinant protein had no transcription activity of reporter genes or toxicity to the yeast cells Y2HGold. Therefore, the pGBKT7-TRAP-A plasmid may be used for the screening of the interactive proteins from Hyalomma anatolicum anatolicum in yeast two-hybrid system.关键词
酵母双杂交系统/环形泰勒虫/TRAP-A蛋白/自激活/细胞毒性Key words
Yeast two-hybrid system/Theileria annulata/TRAP-A Protein/autoactiviation/toxicity分类
农业科技引用本文复制引用
田占成,独军政,高闪电,杜晓悦,于瑞明,刘光远,罗建勋,殷宏..环形泰勒虫TRAP-A蛋白酵母双杂交诱饵质粒的构建与鉴定[J].中国动物传染病学报,2018,26(1):60-65,6.基金项目
国家自然科学基金(31201899) （31201899）
