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稻瘟病新抗性基因Pi39候选基因Pi39E长片段载体的构建

Long Fragment Vector Construction of Pi39E, a Candidate Gene of Novel Blast Resistance Gene Pi39 in Rice
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摘要 稻瘟病新抗性基因Pi39来源于粳稻品种Q15,定位于水稻第12号染色体,前期研究发现候选基因区域预测有12个抗病相关候选基因,其中候选基因Pi39E最大,克隆的难度较大。为了构建Pi39E长片段载体,本实验通过加大反应底物的浓度,目的片段的质量浓度为124 ng/μL,质粒载体的质量浓度为158 ng/μL,目的片断与质粒DNA按1.15∶1的摩尔浓度比进行连接反应,在9μL的总体系中加入1μL的PEG4000,同时采用电击大肠杆菌DH5α转化方法优化实验方案,成功构建该载体。本结果对新抗性基因Pi39精细定位及其功能的研究具有重要意义。 The new rice blast resistance gene Pi39 originates from indica rice cultivar Q15 and is located on chromosome 12 of rice.Previous studies have found that there are 12 candidate genes related to disease resistance in candidate gene regions,among which Pi39 E is the largest candidate gene,and the cloning is more difficult.To construct the Pi39 E long fragment vector,the concentration of the reaction substrate was increased in this experiment.The target fragment mass concentration was 124 ng/μL,the plasmid vector mass concentration was 158 ng/μL,and the ligation reaction between the target fragment and plasmid DNA was carried out at the molar concentration ratio of 1.15∶1.A total of 9μL of the total system was added with 1μL of PEG4000.At the same time,E.coli DH5αtransformation method was used to optimize the experimental protocol and the vector was successfully constructed.This study was of great significance to the fine mapping and function of the new resistance gene Pi39.
作者 陈佳莹 宋斯敏 王春台 刘新琼 Chen Jiaying;Song Simin;Wang Chuntai;Liu Xinqiong(Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China,Key Lab for Biotechnology of state Ethnic Affairs Commission,College of Life Science,South-Central University for Nationalities,Wuhan,430074)
出处 《分子植物育种》 CAS CSCD 北大核心 2019年第9期2882-2885,共4页 Molecular Plant Breeding
基金 国家自然科学基金面上项目(3137306) 中南民族大学中央高校基本科研业务费专项资金(CZP17052)共同资助
关键词 稻瘟病抗性基因Pi39 候选基因Pi39E 载体构建 Rice blast resistance gene Pi39 Candidate gene Pi39E Vector construction
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