Construct GIT1-WT And GIT1-Y293F Lentivirus Vectors And To Investigate The Role Of GIT1-Y293F In Osteoblasts Migration

Objective: To construct GIT1-WT(full-length avian GIT1) and GIT1-Y293F lentivirus vectors and to investigate the role of GIT1-Y293F in osteoblast migration.Methods:The GIT1-WT gene was obtained by polymerase chain teaction(PCR) from mouse gene bank. The PCR products were inserted into PLJM-GFP plasmid. The PLJM-GFP-GIT1-Y293F was obtained by used TaKaRa MutanBEST Kit. Both PLJM-GFP-GIT1-WT and PLJM-GFP-GIT1-Y293F were evaluated by PCR and sequencing. The virus obtained from transfected 293T cells was infected into osteoblasts. The role of GIT1-Y293F in ostroblasts migration was determined by wound healing assay.Results: The PLJM-GFP-GIT1-WT and PLJM-GFP-GIT1-Y293F were constructed successfully. The results of wound healing assay showed that GIT1-Y293F lentivirus vectors can inhibited osteoblast migration.Conclusion: GIT1-Y293F inhibitis ostroblasts migration, which indicate that the tyrosine site 293 of GIT1 is important for GIT1 function in cell migration.