红鳍东方鲀foxo1基因的原核表达及纯化-湖南农业大学学报欢迎您！.pdfVIP

() 2016, 42(1):81–84. DOI:10. 13331/ki.jhau.2016.01.016 Journal of Hunan Agricultural University (Natural Sciences) 红鳍东方鲀 FoxO1 基因的原核表达及纯化 a a a b* ( a.b. 453003) 摘 要鲀RNART–PCR 鲀FoxO1 ORF pET–32/FoxO1Rosetta (DE3)IPTG FoxO1 pET32a/FoxO1 IPTG His 110 000 关 键 词鲀FoxO1 中图分类号S917 Q786 文献标志码：A 文章编号：1007− 1032(2016)01−0081−04 Prokaryotic expression and purification of FoxO1 gene from torafugu Takifugu rubripes a a a b* Zhang Wei Li Cuiping Yang Zhijun He Guangjie (a.School of Basic Medical Science; b.Department of Forensic Medicine, Xinxiang Medical University, Xinxiang, Henan 453003, China) Abstract: The ORF of FoxO1 gene was amplified by reverse transcription-polymerase chain reaction (RT–PCR) from total RNA extracted from adipose tissues of torafugu Takifugu rubripes. PET–32a/Sirt1the prokaryotic expression vectorwas constructed and transformed into Escherichia coli Rosetta (DE3) for FoxO1 expression. Through IPTG inducting expressionthe induced fusion protein was purified and verified by SDS–PAGE and Western Blot. As a result the full-length open reading frame of torafugu FoxO1 cDNA was successfully clonedand the recombinant vector was constructed. The expressed fusion proteins induced by IPTG were soluble protein. The fusion proteins with tag His were also purifiedand the molecular weight of the recombinant protein was about 110 000. Keywords: Takifugu rubripes; FoxO1 gene; prokaryotic expression; protein purification [3–4] [5] [6]