华木莲SgMLPK基因的克隆与亚细胞定位  

Molecular cloning and subcellular localization of SgMLPK gene in Sinomanglietia glauca

作　　者：邓凌帆 邱珊姗 刘苗苗 张建强[1] 王颜波 刘齐元[3] 王建革[1] DENG Lingfan;QIU Shanshan;LIU Miaomiao;ZHANG Jianqiang;WANG Yanbo;LIU Qiyuan;WANG Jiange(College of Forestry,Jiangxi Agricultural University,Nanchang,Jiangxi 330045,China;College of Water Conservancy and Ecological Engineering,Nanchang Institute of Technology,Nanchang,Jiangxi 330029,China;School of Agricultural Sciences,Jiangxi Agricultural University,Nanchang,Jiangxi 330045,China)

机构地区：[1]江西农业大学林学院,江西南昌330045 [2]南昌工程学院水利与生态工程学院,江西南昌330029 [3]江西农业大学农学院,江西南昌330045

出　　处：《福建农林大学学报（自然科学版）》2022年第2期210-216,共7页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(31960418)。

摘　　要：以华木莲心皮为材料克隆了SgMLPK,对其编码蛋白理化性质进行分析,对其结构域、疏水区、跨膜结构、亚细胞定位进行了推断,基于最大似然法构建了系统发育树.将SgMLPK与GFP基因构建了重组表达载体并转化了农杆菌,直接注射到烟草叶片中,36 h后用荧光显微镜观察SgMLPK蛋白的亚细胞定位.结果表明,SgMLPK蛋白定位于细胞质和细胞膜上,共417个氨基酸(碱基1254 bp),具有苏氨酸激酶结构域和信号肽,存在疏水区,无跨膜区域.The SgMLPK gene was cloned and sequenced from the carpel of Sinomanglietia glauca.Based on the sequence of the SgMLPK gene,the motifs,hydrophobicity,signal peptide transmembrane structure,and subcellular localization of SgMLPK protein were predicted.In addition,the phylogenetic tree of SgMLPK protein was constructed by maximum likelihood method.For the subcellular localization of SgMLPK,the recombinant expression vector of SgMLPK and GFP gene was injected into tobacco leaves after successfully being transformed into Agrobacterium tumefaciens.Thirty six hours after the injection,the subcellular location of SgMLPK was observed in the leaf cell of tobacco under fluorescence microscope.The results indicated that the SgMLPK was 1254 bp in length,and coded 417 amino acids.It consisted of 1 motif(Pkinase Tyr),a signal peptide and hydrophobic domain,but had no transmembrane domain.Confocal fluorescence microscopy showed that the SgMLPK protein was localized in the cytoplasm and on the cell membrane.

关 键 词：华木莲 自交不亲和 M位点受体激酶 克隆 亚细胞定位 

分 类 号：Q785[生物学—分子生物学] Q786