Establishment of Transgenic Mice for Expression of Human Protein C Gene in Mamma ry Gland

Objective To establish transgenic mice for the expression of human protein C gene in mamma ry gland. Methods A recombinant plasmid for expression of human protein C(hPC) under the control o f whey acid protein(WAP) promoter was constructed by stepwise cloning method and named as pWPC.The constructed plasmid was digested with PvuⅠ+SmaⅠ,and a 4.6 kb fragment(WAP-hPC-PolyA) was recovered and injected into male pro-nuclei of zygotes of Kunming mice by microinjection.Of the 810 zygotes injected,640 survi val ones were transplanted to 28 mice with pseudopregnancy.A total of 81 offspri ng were born from 18 pregnant mice.The genomic DNAs were extracted from the tail s of offspring for amplification by PCR. Results The genomic DNAs of hPC were amplified from 34 of the 81 offspring.Southern blot proved that the genomic DNAs of hPC were integrated to the genomic DNAs of 20 o f the 34 offspring.On the 7 th day after delivery,ELISA showed the express ion of hPC in the milk of 7 of the 20 mice with hPC DNA integrated. Conclusion The transgenic mice for expression of hPC in mammary gland was successfully esta blished.It laid a foundation of expression of hPC in mammary gland of mice.