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- W2374896098 abstract "Objective To construct eukaryotic expression vector pcDNA3.1(+)/BARF1-his with recombinant of BARF1 gene encoded by EBV and pcDNA3.1(+)-his.Methods The specific primers were designed according to the BARF1 gene cDNA sequence in GenBank provided and restriction sites added at both ends;B95-8 cells was cultured,the cell RNA was extracted,BARF1 gene was amplified by RT-PCR.The PCR products of BARF1 were cloned into pUM-T vector and transformed into E.coli DH5α.The positive clones were screened by ampicillin resistance,identified by restriction enzyme digestion of EcoRI and XbaI.Confirmation the BARF1 gene has been cloned into pUM-T vector.BARF1 fragments were recovered by Gel double-digestion product,and then inserted into the eukaryotic expression vector pcDNA3.1(+)-his,transformed into E.coli DH5α.The positive clones were selected by ampicillin resistance,and insertion direction was confirmed by double digestion and sequencing analysis.Results 666bp products of BARF1 were obtained with cDNA by RT-PCR of B95-8 cell RNA.BARF1 gene fragment was inserted into pUM-T vector by identification of double-digestion.BARF1 gene has been properly connected to the pcDNA3.1(+)-his eukaryotic expression vector through sequencing analysis.Conclusion The eukaryotic expression vector pcDNA3.1(+)/BARF1-his was constructed successfully. BARF1.Prokaryotic vector.Recombinant" @default.
- W2374896098 created "2016-06-24" @default.
- W2374896098 creator A5053117944 @default.
- W2374896098 date "2012-01-01" @default.
- W2374896098 modified "2023-09-26" @default.
- W2374896098 title "Construction eukaryotic vector for BARF1 gene of epstein-barr virus" @default.
- W2374896098 hasPublicationYear "2012" @default.
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