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- W2379370880 abstract "Objective: To construct the bicistronic eukaryotic expression recombinant plasmid pIRSES2-EGFP-hIGF-1 by gene engineering and to investigate its expression in human embryo kidney 293 cells(HEK293).Methods: hIGF-1 cDNA was colned from liver cDNA library by polymerase chain reaction(PCR) and inserted into pMD18-T(TA Clone method).The resultant plasmid was transfected into Escherichia coli TG-1 for amplification.After the DNA sequenced,XhoI and EcoRI were used to cut off the target gene and then inserted into the eukaryotic expression vector pIRSE2-EGFP by the two enzymes.The recombinant plasmid named pIRSES2-EGFP-hIGF-1 was verified by XhoI and EcoRI double-enzyme digestion analysis,and PCR.The plasmid was transfected into HEK293 cells by X-treme GENE HP DNA Transfection reagent in vitro.Then its expression was detection by using fluorescence microscopy and RT-PCR.Results: The hIGF-1 cDNA sequence and insert in recombinant eukaryotic expression plasmid pIRSES2-EGFP-hIGF-1 were both correct.48 hour later,the green fluorescence was observed to be emitted from HEK293 cells after transfection.The RT-PCR result demonstrated that hIGF-1 could express in HEK293 cells after transfected with pIRSES2-EGFP-hIGF-1.Conclusion: The vector pIRSES2-EGFP-hIGF-1 has been constrcuted successfully.The target gene and marker gene can be both expressed in the 293 cell lines,which lays the foundation for further research." @default.
- W2379370880 created "2016-06-24" @default.
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- W2379370880 date "2013-01-01" @default.
- W2379370880 modified "2023-09-22" @default.
- W2379370880 title "Construction and Detection of Bicistronic Eukaryotic Vector pIRSES2-EGFP-hIGF-1" @default.
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