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中文摘要：
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。
英文摘要：
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.
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