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- W2363339156 abstract "Objective:To construct a cDNA subtractive library of genes transactivated by hepatitis B virus(HBV) RNase H protein with suppression subtractive hybridization(SSH) technique and to clone genes associated with its transactivating function.Methods:The mRNA was isolated from HepG2 cells transfected pcDNA3.1(-)-RNase H and pcDNA3.1(-) empty vector,respectively,then cDNA was synthesized,after restriction enzyme RsaⅠ digestion,small sizes cDNA were obtained.Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor.After tester cDNA was hybridized with driver cDNA twice and undergone nested PCR twice and then was subcloned into T/A plasmid vectors to set up the subtractive library.Amplification of the library was carried out with E.coli strain JM109.The cDNA was sequenced and analyzed in GenBank with Blast search after PCR.One clony with no homology with identified genes was primarily confirmed by use of Kozak regulation and the presentation of polyadenyl signal sequence.The new gene was amplified by the reverse transcription PCR(RT-PCR) technique and confirmed with sequencing assay.Results:The new gene was named as DNAPTP4 and enrolled in GenBank,the registered number is AY450392 which consists of 828 nucleotides and codes 276 amino acids.Conclusion:Success in the cloning and identification of DNAPTP4 transactivated by hepatitis B virus RNase H protein by suppression subtractive hybridization provides theoretical basis and research method for the molecular biological mechanism of the transactivation effect by RNase H protein." @default.
- W2363339156 created "2016-06-24" @default.
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- W2363339156 date "2004-01-01" @default.
- W2363339156 modified "2023-09-23" @default.
- W2363339156 title "Identification and Cloning of Human Gene DNAPTP4 Transactivated by RNase H Protein of Hepatitis B Virus" @default.
- W2363339156 hasPublicationYear "2004" @default.
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