title: [Construction and expression of fusion protein TRX-hJagged1 in E.coli BL21].
abstract: This study was purposed to construct prokaryotic expression vector and to investigate the expression of Notch ligand Jagged1 in E.coli. An expression vector pET-hJagged1 was constructed, which can be inserted in Jagged1 with different lengths, but the DSL domain of human Jagged1 should be contained. Then the recombinant plasmids were transformed into the competent cell of E.coli BL21, and the expression of the fusion protein was induced by IPTG. Fusion protein was purified from the supernatant of cell lysates via the Nickel affinity chromatography. The results showed that prokaryotic expression vectors pET-hJagged1 (Bgl II), pET-hJagged1 (Hind I) and pET-hJagged1 (Stu I) were successfully constructed, but only pET-hJagged1 (Stu I) could express the soluble TRX-hJagged1. The purified TRX-Jagged1 protein could be obtained via the Nickel affinity chromatography, and then confirmed by Western Blot. It is concluded that prokaryotic expression vector pET-hJagged1 is successfully constructed, but only pET-hJagged1 (Stu I) can express the soluble TRX-hJagged1 and the TRX-Jagged1 fusion protein is obtained through the prokaryotic expression system, which laid a solid foundation for further to explore the effects of Jagged1 in hematopoietic and lymphoid system.
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Guo-Hui
authors.initials: GH
authors.lastname: Li
authors.affiliation: Department of Traditional Chinese Medicine, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Yu-Zhen
authors.initials: YZ
authors.lastname: Fan
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Si-Yong
authors.initials: SY
authors.lastname: Huang
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Yin
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Ren-An
authors.initials: RA
authors.lastname: Chen
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
authors.forename: Miao-Wang
authors.initials: MW
authors.lastname: Hao
authors.affiliation: Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi. E-mail: liangym@fmmu.edu.cn.
authors.forename: Ying-Min
authors.initials: YM
authors.lastname: Liang
mesh_terms: D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D000072100:Jagged-1 Protein; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D000072099:Serrate-Jagged Proteins
chemical_list: D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D000072099:Serrate-Jagged Proteins