Construction of recombinant plasmid pEGFP-NGB and its expression in cultured neuroglial cells

AIM: To establish recombination plasmid pEGFP-NGB and to investigate the expression of pEGFP-NGB in culture neuroglia cells. METHODS: The NGB cds was isolated by using RT-PCR method with total RNA extracted from fetal Kunming mouse brain, then the NGB cds was cloned into the eukaryotic expression vector pEGFP-C1 of EGFP reported green fluorescence protein. The expression vector of recombinant plasmid pEGFP-NGB was successfully constructed. GeneJamer transfection reagent was used to transfer recombinant plasmid pEGFP-NGB into culture neuroglial cells. The mRNA and protein expression of pEGFP-NGB in culture neuroglial cells were investigated. RESULTS: The positive clone sequencing was consistent with the sequence of Genbank. The NGB mRNA and protein expression of pEGFP-NGB in culture neuroglial cells were detected at high levels. The high expression of green fluorescence protein was observed by fluorescence microscope in culture neuroglial cells. CONCLUSION: The expression vector of recombinant plasmid pEGFP-NGB was successfully constructed and green fluorescence protein was expressed in cultured neuroglial cells.