Construction Of Eukaryotic Vector Of PCV2 And Expression In Dairy Goat Fetal Fibroblasts

The inability and high expenses of bacterial and animal cells for producing complex recombinant proteins make transgenic animals a better choice. Transgenic animal bioreactors can produce more protein production with higher protein activities while at lower costs in mammary milk or urine. All of that make transgenic animal bioreactors a promising technology. This technology opened a new way to make commercial vaccine and other pharmaceutical proteins.Porcine circovirus 2(PCV2) is the main cause of postweaning multisystemic wasting syndrome(PMWS) and rendered great economic losses to the pig industry all over the world. So the antigenic and immune research of PCV2 has been rising these years, which laid a solid foundation for the precaution and treatment of PCV2 infection.In this paper, we constructed the eukaryotic expression vector of PCV2 and then transfected the goat embryonic fibroblast cells and established the positive cell line after G418 screening. The work was supposed to be used as the nuclear donor for the coming somatic cell nuclear transfer procedure.First, PCV2 gene was obtained through RT-PCR, and cloned to the eukaryotic expression vector pc DNA3.1 to make the pc DNA3.1-PCV2 vector. We confirmed the insertion of PCV2 with double restriction enzymes digestion. Then the pc DNA3.1-PCV2 vector was transfected to the goat embryonic fibroblast cells, after G418 screening, the transfected cells were collected to extract RNA and total protein.Real time-PCR and western blotting were performed to examine the expression of PCV2.The results were as follows:products of double restriction enzymes digestion was as expected; the remain cells of G418 screening was established as cell line; PCV2 was expressed according to the results of real time-PCR and western blotting. Our work provided references for the PCV2 products with goat mammary gland bioreactors.