The Primary Study Of T-PAm Goat Mammary Gland Bioreactor Made By Nuclear Transfer

Human tissue type plasminogen activator(t-PA) is a perfect thrombus-dissolved medicament that can dissolve thrombus specifically and effectively but do not induce systemic haemorrhage. The optimal way that cure acute miocardial infarction is treated with t-PA. At the present time, t-PA is mostly produced by culturing mammalian cell and the price is very high. Mammary gland bioreactor is a good mode to produce genie medicament.We constructed the We constructed the mammary gland expression vector pGBCt-PAm, which including 4.1kb goat p-casein promoter and 2.4kb Chicken B-globin insulator in front of the promoter in order to weaken or eliminate "position effect". The availability of the vector was testified through temporal expression in the mammary gland of the femal rabbit.The goat fetal fibroblast cells were transfected by linear GBCt-PAm with electroporation or lipidosome, and after the identification of PCR, we got 7 lines of progenitive cells that examined positive.The two cell line (M9, M10) had been used for the somatic nuclear transfer for the first time. The efficiency of the nuclear transfer was 63.6% (63/99) .Forty-seven clone embryos were transferred into 5 recipient goats. After embryo transfer, two recipient goats have not been seen oestrous within 2 oestrum, and one of the two recipient goats was examined positive by gravid testing, which testified the clone embryos were viable in the uterus of recipient goats.