title: [The recombinant monocyte chemotactic protein 1 expressed in HEK293T cells promotes migration of macrophages].
abstract: OBJECTIVE
  To construct the eukaryotic expression vector of monocyte chemokine protein 1 (MCP-1) driven by the cytomegalovirus (CMV) promoter, transfect the vector into HEK293T cells, and detect its chemotaxis to macrophages.
  
  
  METHODS
  MCP-1 promoter was obtained from mouse genome by PCR, and inserted into the vector named pFLAG-CMV1. We validated it by double-enzymes digestion and sequencing. Then HEK293T cells were transfected with pFLAG-CMV1-MCP-1. The expression of MCP-1 was detected by Western blotting. Finally, we identified the chemotaxis of the recombinant vector to macrophage by Transwell(TM) assay.
  
  
  RESULTS
  The recombinant vector could generate target fragment by double enzyme digestion. HEK293T cells expressed MCP-1 after they were transfected with the recombinant vector, which increased the migration of macrophages.
  
  
  CONCLUSIONS
  MCP-1 expressed by HEK293T cells could apparently increase the migration of macrophages.
authors.affiliation: Department of Nephrology, Xijing Hospital, Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Junlong
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Nephrology, Xijing Hospital, Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Yali
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Han
authors.affiliation: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Qin
authors.affiliation: Department of Nephrology, Xijing Hospital, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China. *Corresponding author, E-mail: liuxiaow@fmmu.edu.cn.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D018932:Chemokine CCL2 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000276:immunology*; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
chemical_list: D018932:Chemokine CCL2; D011993:Recombinant Fusion Proteins