Title: Construction of Eukaryotic Expression Recombinant of NPCEDRG Controlled by Tetracycline(Tet)-Inducible System
Abstract: Objective To construct an inducible eukaryotic expression vector pRevTRE-NPCEDRG containing the open reading frame(ORF) of the NPCEDRG gene for regulating the gene expression by Tet system. Methods The ORF of NPCEDRG gene was amplified from the recombinant plasmid of pcDNA3.1-NPCEDRG by polymerase chain reaction(PCR),and subcloned into the pRevTRE vector after digestion with restriction enzymes BamHⅠand Hind Ⅲ.Then the products were transferred into E.coli DH5α,and the positive clones were screened after being identifid with PCR,restrictive enzymes and sequence analysis. Results The target segment of NPCEDRG was obtained successfully 530 bp in PCR products and 520 bp in digestion ones.The DNA sequence after being sequenced was consistent with the cDNA sequence of NPCEDRG from GeneBank. Conclusion The recombinant plasmid of pRevTRE-NPCEDRG was constructed successfully.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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