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- W3030272323 abstract "Objective To construct an eukaryotic expression vector of human Par-4 gene with green fluorescent protein gene which iS named pIRES2-EGFP/Par-4 and transfect jt into K562 cell line. Methods Using pDNR/Par-4 plasmid as a template.the full length Par-4 cDNA was amplified by PCR and subsequently cloned into T-A vector.Then subcloned into pIRES2-EGFP vector.After identified by digestion of restriefive endonucleases.pIRES2-EGFP/Par-4 was further confirmed by sequencing.Then it was transfected into K562 cells with Superfect reagents.The total proteins were isolated and Par-4 was detected by Western blotting. Results The exact sequences of pIRES2-EGFP/Par-4 vector were confirmed by digestion of restrictive endonucleases and sequencing.After transfection,the expressions of green fluorescent protein were present.The protein expression of Par-4 has been detected in transfected ceils hv Western blotting.Conclusion The vector pIRES2-EGFP/Par-4 has been constructed and could Successfully express Par-4 gene in K562 cells.Key words: Par-4 gene; Eukaryotic expression vector,Transfection; K562 cell" @default.
- W3030272323 created "2020-06-05" @default.
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- W3030272323 date "2009-01-25" @default.
- W3030272323 modified "2023-09-23" @default.
- W3030272323 title "Construction of human Par-4 eukaryotic expression vector and expression in K562 cells" @default.
- W3030272323 doi "https://doi.org/10.3760/cma.j.issn.1009-9921.2009.01.004" @default.
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