Construction And Cell Expression Of Mammary Gland Expression Vector With Position-independent Manner

The purpose of this paper was to construct mammary gland expression vector with position-independent manner, which can protect transgenes from genomic position effects and express independently. The endogenousβ-Casein had been analyzed by RT-PCR( reverse transcription-polymerase chain reaction ) for the assessment of the function of mammary epithelial cells(MEC) and the efficacy of hormonal induction in vitro.Optimized vector pWHERE2v.01 was reconstructed by excising LacZ gene firstly and thereafter the BLG-VP2 gene was inserted in multiple clone site(MCS) flanking by insulators. Thus a mammary gland expression vector that tried to protect transgenes from genomic position effects was constructed.Goat mammary epithelial cells (GMEC) were isolated from lactating goat mammary tissue. RT-PCR and immunochemical method were used to detectβ- Casein expression. The transcription of two mammary epithelial cell lines (C127 and T47D) was also analyzed to compare and assess theβ-Casein gene expression in different mammary epithelial cell lines. The results showed thatβ-Casein gene expression can be detected in primary cultured goat mammary epithelial cells (GMEC-1) derived from lactating goat as well as consequently passaged cells GMEC-3 with the presence of hormones including prolactin, insulin and hydrocortisone.