山东农业科学
山東農業科學
산동농업과학
SHANGDONG AGRICULTURAL SCIENCES
2015年
1期
1-5
,共5页
赵丽%张国嘉%郑世刚%李臻%王庆国%潘教文%刘炜
趙麗%張國嘉%鄭世剛%李臻%王慶國%潘教文%劉煒
조려%장국가%정세강%리진%왕경국%반교문%류위
类受体蛋白激酶%OsESG1%原核表达
類受體蛋白激酶%OsESG1%原覈錶達
류수체단백격매%OsESG1%원핵표체
Receptor-like kinase%OsESG1%Prokaryotic expression
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。
水稻類受體蛋白激酶OsESG1在種胚中特異錶達,屬絲氨痠/囌氨痠蛋白激酶。 OsESG1基因全長cDNA經XhoⅠ、BamHⅠ雙酶切,與同樣經雙酶切的原覈錶達載體pET-28a(+)經T4 DNA連接酶連接,併轉化大腸桿菌感受態細胞BL21。原覈錶達結果顯示,OsESG1可受1 mmol/L IPTG誘導錶達,且蛋白錶達量隨誘導時間的延長而逐漸增加,誘導7 h後,蛋白錶達至較高水平。對不同濃度IPTG誘導下的蛋白錶達情況進行研究,結果顯示,0.1 mmol/L IPTG對OsESG1的誘導錶達效果最好。
수도류수체단백격매OsESG1재충배중특이표체,속사안산/소안산단백격매。 OsESG1기인전장cDNA경XhoⅠ、BamHⅠ쌍매절,여동양경쌍매절적원핵표체재체pET-28a(+)경T4 DNA련접매련접,병전화대장간균감수태세포BL21。원핵표체결과현시,OsESG1가수1 mmol/L IPTG유도표체,차단백표체량수유도시간적연장이축점증가,유도7 h후,단백표체지교고수평。대불동농도IPTG유도하적단백표체정황진행연구,결과현시,0.1 mmol/L IPTG대OsESG1적유도표체효과최호。
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.