山东农业科学2018，Vol.50Issue(5)：7-13,7.DOI:10.14083/j.issn.1001-4942.2018.05.002
水稻类受体蛋白激酶FTPK1的原核表达及其活性检测
Prokaryotic Expression and Activity Detection of Rice Receptor -Like Protein Kinase FTPK1
摘要
Abstract
Receptor -like protein kinases (RLKs) are receptors of signaling molecules of plants .It can be phosphorylated or dephosphorylated through the intracellular kinase domain , thus regulating the transcrip-tion and expression of downstream target gene , participating in the intracellular signal transduction , and pla-ying a role in plant growth and environmental response .In this study, the cDNA of rice variety Zhonghua 11 was used as template to obtain the full -length sequence of FTPK1 gene by PCR, and the pET -28a -FTPK1 prokaryotic expression vector was constructed .An 88 kD fusion protein could be induced by 1 mmol/L IPTG for 12 hours, indicating that FTPK1 could be induced in E.coli.The purified protein product of FTPK1 could be obtained after purification and dialysis by Ni column .The detection of protein kinase activity showed that FTPK1 could phosphorylate the substrates of His and MBP , which showed that FTPK1 has phosphorylation ac-tivity.This research shed some lights for further preparation of the FTPK 1 specific antibody, and lay solid foundation for analysis the physical and biochemical functions of FTPK 1.关键词
水稻/类受体蛋白激酶/FTPK1/原核表达/蛋白纯化/激酶活力检测Key words
Rice/Receptor -like protein kinase/FTPK1/Prokaryotic expression/Protein purification/Kinase activity detection分类
农业科技引用本文复制引用
李颖秀,李臻,潘教文,王庆国,刘炜..水稻类受体蛋白激酶FTPK1的原核表达及其活性检测[J].山东农业科学,2018,50(5):7-13,7.基金项目
山东省自然科学基金面上项目(2016ZRC02178) （2016ZRC02178）
山东省农业科学院青年英才计划项目(2015-2017,2016-2018) （2015-2017,2016-2018）
山东省农业科学院重大科技成果培育计划项目(2015CGPY10) （2015CGPY10）
国家重点研发计划项目(2016YFD0100903) （2016YFD0100903）
