水稻类受体蛋白激酶 OsESG1的原核表达

摘要

Rice receptor-like kinase OsESG1 belongs to serine/threonine protein kinase, which is spe-cifically expressed in rice embryo.In this research, the full length cDNA of OsESG1 was digested by BamHⅠand XhoⅠ, and then linked with the prokaryotic expression vector pET-28a(+) digested by the same double enzymes with T4 DNA ligase.The constructed prokaryotic expression vector of OsESG1-pET-28a(+) was then transformed into host bacteria BL21 for protein expression analysis.The SDS-PAGE showed that OsESG1 could be induced to express by 1 mmol/L IPTG.The expression quantity increased with the elongation of in-duction time, and reached the maximum level after inducing for 7 h.The induction concentration of IPTG was also studied.The results indicated that 0.1 mmol/L IPTG had the best induction effect.%水稻类受体蛋白激酶OsESG1在种胚中特异表达，属丝氨酸／苏氨酸蛋白激酶。 OsESG1基因全长cDNA经XhoⅠ、BamHⅠ双酶切，与同样经双酶切的原核表达载体pET－28a（＋）经T4 DNA连接酶连接，并转化大肠杆菌感受态细胞BL21。原核表达结果显示，OsESG1可受1 mmol／L IPTG诱导表达，且蛋白表达量随诱导时间的延长而逐渐增加，诱导7 h后，蛋白表达至较高水平。对不同浓度IPTG诱导下的蛋白表达情况进行研究，结果显示，0．1 mmol／L IPTG对OsESG1的诱导表达效果最好。