遇文婧;刘志华;王志英;古丽吉米拉米吉提;黄颖;刁桂萍;

• 1:东北林业大学林学院
• 2:黑龙江省森林保护研究所

摘要(Abstract)：

[目的]构建刺激植物响应蛋白Epl1基因的真核表达载体,筛选多拷贝酵母转化子。[方法]以深绿木霉(Trichoderma atroviride)ACCC30153的cDNA为模板进行PCR获得Epl1基因片段,并将目的片段插入表达载体pPIC9K的相应位置,获得重组表达载体,并将pPIC9K-Epl1转入毕赤酵母(Pichia pastoris)GS115中,并用不同中终浓度的遗传霉素G418筛选多拷贝重组酵母菌株。[结果]对重组载体进行PCR及双酶切检测为阳性;在含有终浓度为2 mg/ml遗传霉素G418的YPD平板上筛选得到7株多拷贝的转化子GS115-Epl1,经PCR鉴定1株转化子呈阳性。[结论]Epl1基因的载体构建及多拷贝酵母转化子筛选为以后大量分离纯化Epl1蛋白并研究其功能奠定基础。

关键词(KeyWords)： 深绿木霉ACCC30153;刺激植物响应蛋白;毕赤酵母

Abstract:

Keywords:

基金项目(Foundation): 哈尔滨市科技局青年科技创新人才项目(2013RFQXJ02)资助;; 中央高校基本科研业务费专项资金项目(2572014BA08)

作者(Author): 遇文婧;刘志华;王志英;古丽吉米拉米吉提;黄颖;刁桂萍;

Email:

DOI: 10.13989/j.cnki.0517-6611.2014.19.015

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