Synthesis Of Sheep Oct4and Construction Of Retroviral Vector With Cloned Sox2

Since the induction of mouse iPS cell lines successed, induced pluripotent stem cell research has become a hot area of today’s medical and biological research, Oct4, Sox2, Klf4, lin28and Nanog and other related factors are also more and more important. But there is no report about induced iPS cell lines by using these factors from sheep. In this study, we cloned sheep Sox2coding sequence by molecular biology and synthesis of the Oct4coding sequence, analysis and predict the characteristics of their associated proteins. And then establish of sheep Sox2gene with a retroviral vector and to obtain the viral supernatant. Use the viral supernatant to infect sheep fibroblasts to observe the infection capacity and its transcription level. The study is helpful to the further study of the sheep iPS cell lines. This study results are as follows.(1) Through biological software finishing sheep Oct4gene coding region of1083bp of the theory of sequences, and synthetic. Analysis of the sheep Oct4gene coding region sequence, the results showed98.2%homology with cattle, and the homology is high with the pigs and mice too; the results of analysis sheep Sox2protein sequence showed99.7%homology with cattle, with pigs, and the homology is high with the pigs and mice too. The sheep Sox2protein contains a nuclear localization motif and zinc finger domain.(2) Obtained a1000bp specific bands of sheep Sox2gene by useing fetal sheep muscle tissue, and tested by PCR and restriction endonuclease digestion. The Sox2gene is sequenced and100%similarity with the homologous sequences in GenBank. Analysis of the sheep Sox2gene coding region sequence, the results showed98.2%homology with cattle, and the homology is high with the pigs and mice too; the results of analysis sheep Sox2protein sequence showed99.7%homology with cattle, with pigs, and the homology is high with the pigs and mice too. The sheep Sox2protein contains a nuclear localization motif and zinc finger domain.(3) Build the retroviral vector named pMXs-Sox2, tested by PCR and restriction endonuclease digestion. Viral supernatant obtained has infection and transcriptional capacity.In summary, we obtained sheep Sox2and Oct4gene coding sequence, and constructed the retroviral vector pMXs-Sox2with infection capacity successfully, The study is helpful to the further study of the sheep iPS cell lines.