Transcription Factor Pax6,Sox2 And Six3 Expression And Interaction Analysis In Xenopus Lens Development And Regeneration

During the development of the eye,including many transcription factors,in which Sox2,Pax6,Six3,Prox1,L-maf and so on are most prominent.They are able to form complex regulatory networks,through synergistic or antagonistic effects act on target genes or organization accurately,regulated the normal development and regeneration process.In this experiment,model animal Xenopus laevis was used as the experimental material,by RT-PCR,construction of recombinant expression plasmid,prokaryotic expression and purification,immunity New Zealand White rabbits methods,successfully acquired rabbit anti Sox2,rabbit anti Pax6,rabbit anti Six3 antibodies.Taken as a foundation,these three transcrioption factors expression of the Xenopus eye development and regeneration process were analysised by Western blot.Experimental results showed that in normal development process,issue 12-26 of Pax6 and Sox2 expression tendency basic consistent.The express peak both occured at about issue 18 and 24.Expression of Sox2 and Pax6 after the issue 26 showed a reverse trend.The express peak of Pax6 appeared again at issue 34,but the Sox2 at issue 30.Expression of Six3 decreased significantly after issue 26.The lens regeneration result indicated that,the express peak of Pax6 appeared at 9th day after lentectomy.expression of Six3 showed a clear downward trend,while the expression level of Sox2 was extremely low.Yeast two hybrid results indicated that only the group of the Yeast which cotransformed with pGBK-T7-six3 and pGAD-T7-sox2,showed a obvious blue.The group cotransformed with pGAD-T7-pax6 and pGBK-T7-sox2 only showed a slight blue.These results suggested that the three transcription factors not only play a role in promoting cell proliferation and differentiation,but also an indispensable factor in promoting the lens protein expression and in the process of fibrosis of the lens.Speculated the expression patterns of the three transcription factors in the process of development and lens regeneration of Xenopus laevis.By yeast two hybrid experiment further verified the interaction relationship between the three.Laid a good foundation for the further study of the molecular mechanism of lens development and regeneration process.