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发明公开
CN109971750A 黑胫病菌效应因子的克隆方法 审中-实审

黑胫病菌效应因子的克隆方法
摘要：
本发明公开了黑胫病菌效应因子的克隆方法，通过设计引物；烟草黑胫病菌基因组DNA的提取；PCR扩增烟草黑胫病菌效应因子基因；PCR产物的回收纯化；大肠杆菌DH5α感受态的制备；扩增产物连接pMDTM 19－T Vector并转化大肠杆菌DH5α；转化大肠杆菌；阳性克隆子筛选及菌液的PCR鉴定；筛选到的阳性克隆子进行测序鉴定，最终得到黑胫病菌效应因子。本发明的有益效果是克隆了烟草黑胫病菌的效应因子并分析其功能。
摘要（英）：

The invention discloses a method of cloning Phytophthora parasitica effector. The method comprises: designing primers; extracting Phytophthora parasitica genome DNA; performing PCR (polymerase chain reaction) amplification on tobacco Phytophthora parasitica effector gene; recycling and purifying the PCR product; preparing Escherichia coli DH5lapha competence; linking the amplification product withpMDTM 19-T Vector, and converting Escherichia coli DH5alpha; converting Escherichia coli; screening a positive clone, and performing PCR identification on a bacterial liquid; subjecting the screenedpositive clone to sequencing identification to obtain the Phytophthora parasitica effector. The method has the advantages that the Phytophthora parasitica effector is cloned and its functionality is analyzed.

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