核转录因子TR3及其缺失突变体在酵母双杂交系统中转录激活作用的检测  

THE ASSAY OF TRANSCRIPTIONAL ACTIVATION EFFECT OF TR3 AND DELETION MUTATION IN THE YEAST TWO HYBRID SYSTEM

作　　者：王新兴[1] 杨志华[1] 刘晓华[1] 郭东升[1] 钱令嘉[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050

出　　处：《中国应用生理学杂志》2008年第4期504-507,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金项目(30570753;30430590;30370586)

摘　　要：目的:检测TR3及其转录活性域缺失突变体在酵母双杂交系统中的转录自激活作用。方法:采用PCR方法扩增TR3全长序列及其缺失突变体,构建酵母双杂交系统的诱饵载体pGBKT7-TR3和pGBKT7-TR3/Δ1～690,将pGBKT7-TR3转化感受态酵母菌AH109后培养于含缺失培养基的平板上,检测β-半乳糖苷酶活性,判定其是否具有转录自激活作用。结果:构建了包含TR3全长序列和TR3/Δ1～690序列的诱饵载体,转化酵母菌AH109后在双缺和三缺培养基上未能使β-半乳糖苷酶活性滤纸片变蓝,说明β-半乳糖苷酶报告基因未被激活。结论:TR3及其转录活性域缺失突变体没有转录自激活作用,可用于酵母双杂交系统。Aim： To assay the transcriptional activation effect of TR3 and it＇s deletion mutation in yeast two hybrid system. Methods： The total length of TR3 and TR3/△1- 690 gene was amplified by PCR method and cloned into pGBKT7 vector. Bait vector of pGBKT7- TR3 and pGBKT7-TR3/△1-- 690 was transformed into AH109 competence yeast. Then self activation of the recombination vector was tested by assay the activity of β-galactosidae. Results： The pGBKT7-TR3 and pGBKT7-TR3/△1- 690 vector was successfullyconstructed. The filter paper containing β-galactosidae didn＇t changed to blue showed that the reporter gene wasn＇t activationed. Conclusion： TR3 and TR3/△1 -690 hadn＇t the activity of transcriptional activation.

关 键 词：TR3 酵母双杂交 转录激活作用 

分 类 号：Q426[生物学—神经生物学]