RESUMO
PURPOSE: To construct a lentiviral eukaryotic expression vector containing two genes of cbfa1 and satb2. METHODS: The aim genes of cbfa1 and satb2 were amplified from plasmids by PCR. After TA cloning, the positive clones were identified by restrictive enzyme digestion and commercial DNA sequencing. Cbfa1 and satb2 with correct sequences were ligated upstream and downstream to pIRES, respectively, to construct pIRES-cbfa1-satb2. Then cbfa1-Ires-satb2 fragment was obtained by double digestion, and inserted into corresponding enzyme cut sites of pLentinTrident1-CMV which had been added resistance gene neo/kana to construct the lentiviral eukaryotic expression vector pLentinTrident1-CMV-cbfa1-Ires-satb2. RESULTS: We amplified the genes cbfa1 and satb2 by PCR and connected them with pLentinTrident1-CMV by internal ribosomal entry site of mediator pIRES successfully. The result was identified by PCR, restrictive enzyme digestion and sequencing. CONCLUSIONS: Recombinant lentiviral eukaryotic expression vector containing both cbfa1 and satb2 genes is successfully constructed. This provides a foundation for further studies on their functions.
