PnsICE1基因酵母双杂交诱饵载体构建及转录自激活检测
作者：王艳敏
来源：《现代农业科技》2015年第13期
摘要 为了更好地研究与PnsICE1基因互作的蛋白，利用向前方PCR方法扩增含有pGBKT7退伍义务兵安置条例载体同源序列的PnsICE1基因片段，构建酵母双杂交载体。测序结果表明，构建的目标基因序列正确。将诱饵载体质粒转化酵母菌株Y2H感受态细胞，PnsICE1转化菌在SD/-Trp营养缺陷型培养基中正常生长，在SD/-Trp/X-a-Gal筛选培养基上长出蓝色的酵母单菌落，说明PnsICE1转录因子具有转录自激活活性。
关键词 PnsICE1；诱饵载体；转录自激活；测序
道路交通安全法实施条例中图分类号 Q943.2 文献标识码 A 文章编号投篮姿势图解 1007-5739（2015）13-0186-02
Construction of Yeast Two-Hybrid Bait Vector of PnsICE1 and Testing of Autonomous Transcriptional Activation
WANG Yan-min 1，2
（设计翻译1 Forestry Science Rearch Institute of Heilongjiang Province，Harbin Heilongjiang 150081新闻阅读； 2 Key Laboratory of Fast-Growing Tree
Cultivating of Heilongjiang Province）
Abstract In order to screen the PnsICE1 interacted proteins，the CDS quence of PnsICE1 contains pGBKT7 vector homologous quences was amplified using PCR and inrted into pGBKT7 vector to construct the bait vector for yeast two-hybrid（Y2H）system. Sequencing result showed that the PnsICE1 quence was inrted into pGBKT7 vector in correct reading frame. Furthermore，the bait vector was transformed into Y2H yeast strain to test the autoactivation. The transformed yeast grew very well on SD/-Trp plate，辞去领导职务in the SD/-Trp/X-a-Gal screening yeast，grown on the medium blue single colony，冰淇淋的图片PnsICE1 transcription factor has the transcriptional activation activity.
