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- W2347574137 abstract "Objective To construct the eukaryotic expression vector of human regucalcin(RGN),and to detect their expressing in human HK-2 cell.Methods The full-length cDNA encoding RGN gene was amplified by reverse transcription polymerase chain reaction(RT-PCR) and subcloned to eukaryotic expression vector pEGFP-N1,the recombinant pEGFP-RGN was transformed into E.coli.DH5α,identified by double digestion with restriction endonucleases and sequencing,then RGN-EGFP were retransformed into pcDNA3.1(+)-zeocin by restriction endonucleases,the recombinant pcDNA3.1-RGN-EGFP-zeo was transformed into E.coli.DH5α,identified by double digestion with restriction endonucleases and sequencing.The sequence of RGN full-length cDNA was confirmed by blasting to Genbank.The recombinant plasmids were transfected into HK-2 cells by lipofectamine method.The expression of RGN mRNA in HK-2 cells was assayed by fluorescence microscopy and laser scanning confocal microscopy(LSCM).Results The sequence of RGN cDNA was corrected by blasting to Genbank.The RGN fusion proteins were expressed in the cells transfected with pcDNA3.1-RGN-EGFP-zeo plasmid.Green fluorescence was observed by fluorescenece microscopy and LSCM on the cells transfeced with pcDNA3.1-RGN-EGFP-zeo plasmid.Conclusion The recombinant eukaryotic expression vector pcDNA3.1-RGN-EGFP-zeo was successfully constructed and expressed in the HK-2 cells." @default.
- W2347574137 created "2016-06-24" @default.
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- W2347574137 date "2006-01-01" @default.
- W2347574137 modified "2023-09-28" @default.
- W2347574137 title "Construction of human regucalcin eukaryotic expression vectors and expression" @default.
- W2347574137 hasPublicationYear "2006" @default.
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