The Isolation And Primary Cell Culture Of Mammary Gland Epithelial Cell From Rabbit's Milk For Activity Detection Of Mammary Gland Specific Promoter

Objective: Based on mammary gland specific expression vector pCA1 containing bovine α-S1-casein transcription regulation sequence, the mammary gland expression vector pCA1-GFPuv is constructed and used to transfect the isolated primary mammary gland epithelial cells from rabbit's milk for detecting activity and mammary gland's tissue specificity of transcription regulation sequence in vector pCAl.Methods: The GFPuv gene as reporter was obtained by using PCR and then it was recombinated into the vector pCA1 to construct mammary gland expression vector pCA1-GFPuv. The isolated rabbit's primary mammary gland epithelial cells were transfected with the vector pCA1-GFPuv by liposome method. Finally the transfected cells were observed under inversed fluorescence microscope to detect activity andmammary gland's tissue specificity of transcription regulation sequence.Results: The construct of expression vector pCA1-GFPuv was confirmed byrestriction enzyme assay and sequencing. After transfected into primary mammarygland epithelial cells, the expression of GFPuv can be observed under inversedfluorescence microscope.Conclusions: The expression vector pCAl-GFPuv has activity and mammary gland'stissue specificity by expressing the GFPuv gene in the rabbit's primary mammarygland epithelial cells.