Constructing eukaryotic expression plasmid pcDNA3-rhHIF-1α of the recombinant human hypoxia-inducible factor-1α

Objective:To construct the eukaryotic expression plasmid pcDNA3-rhHIF1α of recombinant human hypoxia-inducible factor-1α so as to prepare for the clone of human genes and the study of its antibodies.Methods: Total RNA was extracted from Hela cells,and cDNA library was constructed by the reverse transcriptional PCR method.Then the enzymes were inserted into the pcDNA3 vector.Results: Recombinant eukaryotic expression plasmid pcDNA3-rhHIF-1α was gained.The target gene obtained through PCR amplification had the same molecular size as it was expected.Then insert the target gene into the right place of the pcDNA3 vectors after measuring the sequence.Conclusion: The pcDNA3 is a highly expressive eukaryotic vector,and through which the recombinant human HIF-1α can express highly the target gene.The pcDNA3-rhHIF-1α plasmid has been constructed successfully and will lay a certain foundation for the further research in the gene clone and expression of human HIF-1a,and the preparation for the antibody.