2006Journal of OncologyRequires access

Construction and Sequence Analysis of the Transcriptional Small Hairpin RNA Recombinant Plasmid Targeting Bcl-2 Gene

Liu Fang-xin

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Abstract

[Purpose] To study the new approach of tumor gene therapy by constructing recombinant using Bcl-2 as targeting gene with the help of RNA interfering and by conducting sequence analysis. [Method] Two designed DNA sequences with small hairpin structure were cloned into the transcripted carrier Pgenesil-1 in order to construct recombinant, and the recombinant was transformed into DH5a strains.Then the sequence analysis was carried out after the recombinant plasmid was identified by enzyme restriction. [Results] The synthesized DNA fragment was cloned into the carrier after annealing. It was identified as an aim sequence by enzyme restriction and sequence analysis. The recombinant pasmid targeting Bcl-2 gene was successfully constructed and the aim sequence was obtained. [Conclusion] Successfully constructing the recombinant plasmid provides the scientific base for searching a new gene therapy method for cancer.

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What this paper is about

[Purpose] To study the new approach of tumor gene therapy by constructing recombinant using Bcl-2 as targeting gene with the help of RNA interfering and by conducting sequence analysis. [Method] Two designed DNA sequences with small hairpin structure were cloned into the transcripted carrier Pgenesil-1 in order to construct recombinant, and the recombinant was transformed into DH5a strains.Then the sequence analysis was carried out after the recombinant plasmid was identified by enzyme restriction. [Results] The synthesized DNA fragment was cloned into the carrier after annealing. It was identified as an aim sequence by enzyme restriction and sequence analysis. The recombinant pasmid targeting Bcl-2 gene was successfully constructed and the aim sequence was obtained. [Conclusion] Successfully constructing the recombinant plasmid provides the scientific base for searching a new gene therapy method for cancer.

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Available abstract

[Purpose] To study the new approach of tumor gene therapy by constructing recombinant using Bcl-2 as targeting gene with the help of RNA interfering and by conducting sequence analysis. [Method] Two designed DNA sequences with small hairpin structure were cloned into the transcripted carrier Pgenesil-1 in order to construct recombinant, and the recombinant was transformed into DH5a strains.Then the sequence analysis was carried out after the recombinant plasmid was identified by enzyme restriction. [Results] The synthesized DNA fragment was cloned into the carrier after annealing. It was identified as an aim sequence by enzyme restriction and sequence analysis. The recombinant pasmid targeting Bcl-2 gene was successfully constructed and the aim sequence was obtained. [Conclusion] Successfully constructing the recombinant plasmid provides the scientific base for searching a new gene therapy method for cancer.

Key concepts: Recombinant DNA, Gene, Plasmid, Restriction enzyme, Sequence analysis, Small hairpin RNA, Molecular biology, DNA sequencing

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