Technologies

Technical Summary

The pBI-GL-HRE V6R plasmid containing six copies of hypoxic responsive element (HRE) derived from the human VEGF gene was used to generate a plasmid (pACN188) containing an alkaline phosphatase gene under the V6R HRE promoter. LN229 cells were cotransfected with the resulting plasmid and pcDNA3 vector carrying a neomycin-resistant gene. Stable clones were selected by through antibiotic resistance and further tested for hypoxia inducible alkaline phosphatase expression. The clone (LN229-HRE-AP #16) with the highest expression ratio of alkaline phosphatase enzymatic activity under hypoxia versus normoxia was selected for further use.