| Many remarkable progress has been made in mammal somatic nuclear transfer technology(SCNT). After the first cloned “Dolly†was born in 1997, 19 different kinds of mammals has been obtained successfully cloned offspring through somatic nuclear transfer techniques, throughout the world set off a "clone" craze. Sika deer is a very rare animals, it has a close relationship with human life. It can provide more expensive medical care for human medicines, can also provide humans with food and clothing. We are still seeing reports of sika deer in SCNT.But due to the reduced number of sika deer leads to lake of experimentals, the research of sika deer embryos in the stagnant situztion. So looking at alternative substitute of Sika Deer oocytes is deer interspecific cloning research in a noteworthy key problems.This thesis is mainly on procedures for the production of sika deer-bovine interspecific somatic nuclear transfer technology. With the results we found that reprogramming capability of fetal fibroblast cells as donor cells is larger than this of fibroblasts as donor cells, which is more suitable for somatic cell nuclear transfer(13.1% vs. 7.1%). Our results suggested that280 V of electrical activation is most effective for interspecific cloned embryo d evelopment.The blastocyst rates of the interspecies cloned embryos were significantly higher when the embryos were cultured in SDSOF medium than when cultured in SOFAA medium(10.7% vs.5.6%, P<0.05).And at the same time, TSA treatment effects on the development of interspecific somatic nuclear transfer, This thesis is to improve the efficiency of preimplantation embryos by ISCNT. We established a pEGFP-N1 sika deer fibroblast cell line, treated donor cells and cloned embryos with several TSA concentrations.We found that the H4K5 protein signal in the pEGFP-N1 fibroblast strain was up-regulated upon TSA treatment in a concentration-dependent manner. The blastocyst rates of interspecies cloned embryos from clone 7 or 11 were significantly higher than from clone 5(9.2% or 10.7% compared with4.3%, P<0.05). In summary, we found that bovine oocytes could support sika deer donor celldedifferentiation and nuclear reprogramming. |
