Effect Of Sox2 Gene On Early Development Of Mongolian Sheep Embryos Cloned From Mesenchymal Stem Cells

Somatic cell clone as the key technique method in the animal genetic breeding system are always trouble in the less efficiency and embryo abnormal issues, about this issues the study are focus on the optimize donor cell and the genetically modified mechanism in the cloned embryos process of reprogram. this research are plan to use the Mongolia sheep trans Sox2 gene’s mesenchymal stem cells as the donor cellconstruct the cloned embryo by the nuclear transfer, through contrast with the control group.analyze the Sox2’s influence on the early stage cloned embryos development offer some new ideas to improve transgenic cloned efficiency, the main research are as follows:1. Successfully cloned the Mongolia sheep Sox2 gene’s cDNA by the PCR method, and construct the Eukaryotic expression vector pSox2-EGFP.2. Use the lipofection method transfect the pSox2-EGFP into the mesenchymal stem cells which are from the Mongolia sheep fetus, and select the 3 positive cell lines, detect by the real-time qPCR, contrast with the control group, the transfect group the Sox2 gene’s expression has the great increase, the expression is 2.27-4.86 times the control group’s.3. Use the positive cell as the donor cell, construct the cloned embryo, the fusion rate is the 82.58% (fusion embryo 166/construct embryo 201); the cleavage rate is 80.72% (cleavage embryo 134/fusion embryo 166); morula rate 31.37%(morula embryo 52/fusion embryo 166and 1 blastula). The develop rate from the transgenic cloned embryo contrast with the general mesenchymal stem cells has no significant difference.