[Construction and expression of a fusion protein containing extracellular domain of human Jagged1 and Fc fragment of human IgG1].
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.
Jia
Hong-xia
HX
Department of Hematology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. jhx121883@sina.com
Zhang
Ping
P
Zhou
Peng
P
Han
Hua
H
Liang
Ying-min
YM
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
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