Cloning and Expression of Pun1 Gene Controlling Pungency of Pepper (Capsicum spp.)  

控制辣椒辣味基因Pun1的克隆及表达（英文）

作　　者：董雨薇[1] 孙樱燃 毕然[1] 孙宁莉 阮文渊[1] 李越[1] 王晶莹[1] 郭庆勋[1] 

机构地区：[1]吉林大学植物科学学院,吉林长春130062

出　　处：《Agricultural Science & Technology》2016年第11期2483-2488,共6页农业科学与技术（英文版）

基　　金：Supported by College Student Innovation Fund Project of Jilin University(2015821243)~~

摘　　要：[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, with a total length of 1 457 bp, coding 440 amino acids. [Result] Phylogenetic analysis showed that Capunl was closest to Pun1 of C. chinense, with a genetic distance of 0.019 3. Plant expression vector pCAM-Punl-GFP was constructed and transformed into to- bacco, and it was found that the protein coded by fusion gene Punl：：GFP was lo- cated on cell membrane. Prokaryotic expression vectors were constructed, and by SDS-PAGE and Western Blot detection, an induced protein with a molecular weight of 63 ku was obtained. It was found by real-time fluorescence quantitative expres- sion that Pun1 gene was expressed at the highest level 30 d after flowering, de- creased then, and could not be detected substantially 40 and 45 d after flowering. [Conclusion] This study provides information and reference for molecular regulation mechanism of Pun1 gene.[目的]对控制辣椒辣味基因pun1的克隆及表达进行研究。[方法]以益都红(Capsicum annuum L)为材料,获得Capun1基因c DNA序列,全长1 457 bp,编码440个氨基酸;研究了其瞬时表达、原核表达和实时荧光定量表达。[结果]系统进化分析显示,CAPUN1与同属C.chinense辣椒PUN1遗传距离最近,为0.019 3。构建植物表达载体p CAM-Pun1-GFP转化烟草瞬时表达,发现融合基因Pun1::GFP编码的蛋白定位在细胞膜上。构建原核表达载体,通过SDS-PAGE和Western Blot检测,得到了一个分子量为63 ku的诱导蛋白。实时荧光定量表达发现Pun1基因在花后30 d表达量最高,而后开始下降,花后40和45 d基本不表达。[结论]该研究为研究辣椒辣味基因Pun1的调控分子机制提供了信息和参考。

关 键 词：PUNGENCY Pun1 gene Transient expression Prokaryotic expression 

分 类 号：S641.3[农业科学—蔬菜学]