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.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
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)
关键词
TR3
酵母双杂交
转录激活作用
TR3
yeast two hybrid system
transcriptional activation effect
