Objective     To construct the recombinant adenovirus encoding mouse wild type FGFR3 cDNA.      Methods     Mouse FGFR3 cDNA obtained from MoFR3/SV was subcloned into plasmid pBluescript KS and further cloned into plasmid pAdTrack-CMV. The plasmid (pAdTR3) was transferred into BJ5183 cells which contained the adenovirus plasmid (pAdeasy-1) to produce recombinant adenovirus vector encoding FGFR3 cDNA (pAdE-FGFR3). The recombinant adenovirus vector was identified and  transfected into the adenoviral packaging cell HEK293 by lipofectamine 2000 to get recombinant adenovirus particles. The adenovirus was confirmed by po

lymerase chain reactin (PCR) and its titer was determined. Then the recombinant vector was transfected into HT29 cells. The expression of FGFR3 mRNA and protein in HT29 cells was assayed  by RT-PCR and Western blotting.      Results     The recombinant adenovirus vector encoding FGFR3 cDNA was correctly constructed and confirmed by restriction endonuclease analysis and DNA sequencing analysis. The transfected HEK 293 cells were lysed by freeze-thawing to obtain the recombinant adenovirus in the lysate. Further

PCR product of the lysate confirmed the presence of recombinant adenovirus. The viral titer was 3×10

9

pfu/ml. RT-PCR and Western blotting showed that pAdE-FGFR3 transfected HT29 group expressed FGFR3 higher than that of GFP controlled group.      Conclusion     Infective recombinant adenovirus encoding FGFR3 cDNA was successfully obtained by plasmid homogenous recombination in bacteria

and highly expressed in HT29 cells after transfection

which paves a way for studying the effect of FGFR3 in bone fracture healing.