Expression Of Human Proinsulin Gene In Cell And Mouse Under The Control Of Both Goat BLG Regulatory Region And CMV Sequence

At present, there is no effective treatment for diabetes except for injecting ectogenesis insulin. Nowadays, with the development and application of the transgenic techniques, the study of mammary gland bioreactor to produce insulin is a focus in life and biotech areas.A goatβ-lactoglobulin(BLG) regulatory sequences, enhancer sequence, mark gene (Neo gene) and human proinsulin genomic DNA were used to construct human proinsulin mammary gland-specific expression vector. This paper studied on the expression of the vector in goat mammary gland epithelial cells and transgenic mice. And we tested the feasibility of producing in mammary gland bioreactor.The mammary epithelial cells were transfected with foreign gene by electroporation. After two weeks selection by 400μg/ml G418, the stably cell clones were obtained. After inducing with prolactin, ELISA detection showed six positive cell strains were obtained.The foreign gene was microinjected into fertilized mice eggs. PCR analysis showed 1 of 43 mice was transgenic. The milk and the blood of the PCR-positive mouse were detected by ELISA. The results indicate that the vector could be mammary gland-specific expressed in mice.In our study, human insulin gene was expressed in the mammary gland epithelial cells and the tissues of mouse mammary glands with mammary gland specifically. These results indicate that the mammary gland-specific expression vector can be used as an expression vector for development of mammary gland bioreactors.