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Cloning, prokaryotic expression, and subcellular localisation of the TaMAPK10-like gene in common wheat

文献类型： 外文期刊

作者： Nie, Yingbin 1 ; Ma, Songmei 3 ; Ji, Wanquan 1 ;

作者机构： 1.Northwest A&F Univ, Coll Agron, State Key Lab Crop Stress Biol Arid Areas, Yangling 712100, Shaanxi, Peoples R China

2.Xinjiang Acad Agrireclamat Sci, Inst Crop Res, Shihezi 832000, Peoples R China

3.Shihezi Univ, Coll Sci, Dept Geog, Shihezi 832000, Peoples R China

关键词： mitogen-activated protein kinase; prokaryotic expression; subcellular localisation

期刊名称：CANADIAN JOURNAL OF PLANT SCIENCE （ 影响因子：1.018； 五年影响因子：1.242 ）

ISSN： 0008-4220

年卷期： 2019 年 99 卷 4 期

页码：

收录情况： SCI

摘要： Mitogen-activated protein kinase (MAPK/MPK) is a group of serine-threonine protein kinases that are activated by different extracellular stimuli. To explore the function of MAPK in wheat infected with powdery mildew, a new wheat germplasm N9134 was used to obtain the full-length MAPK gene and the MAPK sequence was used to identify its prokaryotic expression and subcellular localisation. Wheat MAPK was obtained by homologous gene cloning and designated as TaMAPK10-like. The open reading frame of TaMAPKIO-like was 1638 bp, which coded a deduced protein of 545 amino acids. Phylogenetic analysis revealed that TaMAPK10-like was most closely related to MAPK10-like of Aegilops tauschii at the protein level. It had high nucleotide similarity with the reported MAPK gene in A. tauschii, Sorghum bicolor, and Setaria italica, and had features typical of MAPK family genes. Subcellular localisation showed that TaMAPK10-like was mainly located in the cytoplasm along the microtubules, and a small number were located in the cell membrane and the nucleus. A pMD-MAPK10-like fusion expression vector was constructed and the TaMAPK10-like fusion protein was 70 kDa. The expressions of protein in bacteria were best obtained using 0.5 mmol L-1 isopropyl beta-D-1-thiogalactopyranoside at 37 degrees C for 12 h. These results provide the basic data for further understanding the biological function of the TaMAPK10-like gene.

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