HBD3 mammary gland specific expression vector and constructed recombinant cell
An expression vector and specific technology, applied in the field of genetic engineering, can solve problems such as negative effects, complicated results analysis, environmental safety problems of transgenic animals and plants, etc., and achieve the effect of avoiding potential safety hazards.
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[0035] The HBD3 mammary gland-specific expression vector provided by the present invention first constructs the HBD3 mammary gland-specific expression vector pARNG-HBD3 with the attB sequence, and secondly makes the exogenous expression vector pARNG-HBD3 site Specifically integrated into the genome of bovine fetal fibroblasts, the expression of red fluorescent protein was observed under a fluorescent microscope, and positive cells were obtained through G418 screening, and PCR identification was performed to confirm that the target gene HBD3 was integrated into the genome of bovine fetal fibroblasts; then Positive cell clones were treated with Cre recombinase, and the antibiotic selection marker, red fluorescent protein gene and terminator between the two LoxPs in the same direction were removed, so that the Pcmv IE promoter and EGFP gene were spliced, and the green fluorescent protein was expressed. After flow cytometry The recombinant bovine fetal fibroblasts containing the HB...
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