title: [Construction and expression of a fusion protein containing extracellular domain of human Jagged1 and Fc fragment of human IgG1].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector containing the extracellular domain of human Jagged1 and Fc fragment of human IgG1 fusion gene and to express the hJagged1(ext)-Fc fusion protein in mammalian cells.
  
  
  METHODS
  The extracellular domain of human Jagged1 gene was cloned from normal human bone marrow cells. In order to obtain hJagged1(ext)-Fc fusion gene, the extracellular domain of Jagged1 gene was inserted into pBluescript-sk II-hc gamma 1, and then the fusion gene was inserted into pEF-BOSneo to get the eukaryotic expression vector pEF-BOSneo-hJagged1(ext)-Fc. The recombination plasmid was transiently transfected into COS7 cells and the expression of the fusion protein was identified by RT-PCR, immunofluorescent assay and sandwich ELISA.
  
  
  RESULTS
  The extracellular domain of human Jagged1 gene was effectively amplified. The DNA sequencing result showed that the constructed plasmid containing hJagged1(ext)-Fc fusion gene was the same as designed. The fusion protein was successfully expressed in mammalian cells.
  
  
  CONCLUSIONS
  The hJagged1(ext) gene has been successfully cloned and expressed, which provides a new fusion protein for further experiments, for example, the Jagged1(ext)-Fc fusion protein can be used as a new stimulator for proliferation of hematopoietic stem/progenitor cells in-vitro.
authors.affiliation: Department of Hematology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. jhx121883@sina.com
authors.forename: Hong-xia
authors.initials: HX
authors.lastname: Jia
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Zhou
authors.forename: Hua
authors.initials: H
authors.lastname: Han
authors.forename: Ying-min
authors.initials: YM
authors.lastname: Liang
mesh_terms: D000818:Animals; D019556:COS Cells / Q000378:metabolism; D002135:Calcium-Binding Proteins; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005822:Genetic Vectors; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis* / Q000235:genetics; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D010957:Plasmids; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D000072099:Serrate-Jagged Proteins; D014162:Transfection
chemical_list: D002135:Calcium-Binding Proteins; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D000072099:Serrate-Jagged Proteins