Objective The recombinant adenovirus vector of Racl and its mutants were constructed for study the role racl gene plays in the migration of MSCs. Method The pAdTrack-CMV-Racl was constructed by PCR with pMD-19-T-Racl recombinant plasmid as template, enzyme digestion and ligation. The pAdTrack-CMV-Racl lineared by Pmel was co-transformed into BJ5183 with pAdEasy-1. The pAdEasy-1-pAdTrack-CMV-Racl recombinant adenovirus vector was lineared with Pad and then transfected into the package cell QBI-293A. The Racl and its mutants recombinant adenovirus were obtained and the virus multiplicity of infection ( MOI) of mesenchymal stem cells were identified after the detection of the virus titer. Results The Racl and its mutants were confirmed by sequencing, and about 107 pfu/ml of high titer recombinant virus was successfully packaged after 2-3 amplification. For mesenchymal stem cells infection, the optimal virus of Ad-Racl was 150MOI. Conclusion The pAdEasy-1-pAdTrack-CMV-Racl, pAdEasy-l-pAdTrack-CMV-RaclQ61L, pAdEasy-1- pAdTrack-CMV-RaclG12V, pAdEasy-1-pAdTrack-CMV-RaclT17N vectors were constructed and the Racl and its mutants recombinant adenovirus ( Ad-Racl, Ad-RaclQ61L, Ad-RaclG12V, Ad-RaclT17N) were obtained successfully.
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