Adenovirus shuttle plasmid pShuttle CMV-+ VEGF121-IRES-hrGFP-1 Construction and identification of.docVIP

 PAGE \* MERGEFORMAT 10 Adenovirus shuttle plasmid pShuttle CMV-+ VEGF121-IRES-hrGFP-1 Construction and identification of [Abstract] Objective To construct the adenovirus shuttle plasmid pShuttle CMV-+ VEGF121-IRES-hrGFP-1, for building epitope tag expression of vascular endothelial growth factor 121 (vascular endothelial growth factor 121, VEGF121), and the simultaneous expression of green fluorescent protein (green fluorescent protein, GFP) reporter molecule in the eukaryotic expression adenovirus vector basis. Methods for Plasmid gene carried pTG19T-VEGF121 VEGF121 gene sequencing and sequence within the existing restriction enzyme recognition sites for analysis, using PCR (Polymerase Chain Reaction, PCR) technique pTG19T-VEGF121 VEGF121 gene mutations carried to remove after the translation termination codon and the sequence of the gene sequence before and after adding a new NotI and XhoI restriction sites. will The VEGF121 gene mutation (+ VEGF121) directed into the adenovirus shuttle plasmid with pShuttle CMV-IRES-hrGFP-1, obtained by electrophoresis and sequencing of recombinant plasmid. Results The recombinant plasmids by electrophoresis and sequencing for the correct clone. Conclusion The construction of the pShuttle CMV-+ VEGF121-IRES-hrGFP-1. [Keywords:] adenovirus shuttle vector; vascular endothelial growth factor 121 Abstract: Objective To clone vascular endothelial growth factor 121 (VEGF121) gene into the adenovirus shuttle plasmid pShuttleCMV-IRES-hrGFP-1, preparing for construction of a novel recombinant adenovirus vector expressing the VEGF121 fused to FLAG epitope and humanized Renilla reniformis green fluorescent protein 1 (hrGFP-1) as a reporter on the same transcript. Methods The VEGF121 gene contained in the plasimid of pTG19T-VEGF121 was sequenced, and the profile of restriction endonuclease sits existing in the sequence was analysed. Then the translation stop codon TAG was removed. Meanwhile, NotI and Xho I restriction sites we