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2009•Chinese Journal of Birth Health & HeredityRequires access

Construction of eukaryotic recombinant expression vector pcDNA3.1 with rat VEGF120 gene

Ji Won Hong

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Abstract

Objective:To construct recombinant eukaryotic expression vector pCDNA 3.1 with rat VEGF120 gene.Methods:Total RNA was extracted from rat Hypoxia brain.The VEGF120 gene was amplified by RT-PCR,then was inserted reversely into the eukaryotic expression vector pcDNA3.1after digestion and ligation using restriction endonucleases and ligase.The recombinant plasmid was sequenced and transformed into E.Coli JM109.Results:The positive clones were acquired after being identi-fid with restrictive enzymes and sequence analysis.After being sequenced with DNA auto-sequence analysis instrument,The VEGF129 DNA sequence of rat was searched for alignment with NCBI Blast program.Conclusion:The recombinant eukaryotic expression vector with rat VEGF120 gene was obtained successfully.

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What this paper is about

Objective:To construct recombinant eukaryotic expression vector pCDNA 3.1 with rat VEGF120 gene.Methods:Total RNA was extracted from rat Hypoxia brain.The VEGF120 gene was amplified by RT-PCR,then was inserted reversely into the eukaryotic expression vector pcDNA3.1after digestion and ligation using restriction endonucleases and ligase.The recombinant plasmid was sequenced and transformed into E.Coli JM109.Results:The positive clones were acquired after being identi-fid with restrictive enzymes and sequence analysis.After being sequenced with DNA auto-sequence analysis instrument,The VEGF129 DNA sequence of rat was searched for alignment with NCBI Blast program.Conclusion:The recombinant eukaryotic expression vector with rat VEGF120 gene was obtained successfully.

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Available abstract

Objective:To construct recombinant eukaryotic expression vector pCDNA 3.1 with rat VEGF120 gene.Methods:Total RNA was extracted from rat Hypoxia brain.The VEGF120 gene was amplified by RT-PCR,then was inserted reversely into the eukaryotic expression vector pcDNA3.1after digestion and ligation using restriction endonucleases and ligase.The recombinant plasmid was sequenced and transformed into E.Coli JM109.Results:The positive clones were acquired after being identi-fid with restrictive enzymes and sequence analysis.After being sequenced with DNA auto-sequence analysis instrument,The VEGF129 DNA sequence of rat was searched for alignment with NCBI Blast program.Conclusion:The recombinant eukaryotic expression vector with rat VEGF120 gene was obtained successfully.

Key concepts: Recombinant DNA, Gene, Molecular biology, Restriction enzyme, Biology, Plasmid, Vector (molecular biology), Expression vector

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