Paired box 6(Pax6)是1段高度保守基因,在调控脊椎动物眼睛发育的过程中起到重要的作用。Paired box 6variant(Pax6variant,VP)是花背蟾蜍体内的Pax6基因家族的同源基因。本实验主要关注非洲爪蟾Pax6基因与花背蟾蜍vp基因之间的异同,两者对细胞增殖、周期调控及细胞命运决定的影响,以及相关位点改变对增殖及其下游的影响,以此来预测具体起主要功能的位点。分别对PAX6和VP 2种蛋白进行磷酸化位点预测及序列比对后发现,在蛋白VP中,不论是可能的磷酸化位点还是可以对其进行磷酸化的激酶识别位点都发生了显著减少,这些都是位于C末端的PST结构域发生的重大改变。针对其不同的磷酸化位点设计的点突变结构,推测该基因的变化是以373位磷酸化位点为主导,其他磷酸化位点对其有相应的影响。
Paired box 6(Pax6)is a highly conserved genes,it plays an important role in the process of regulating the vertebrate eye development.Paired box 6variant(Pax6variant,VP)is a Bufo raddei Pax6 gene families of homologous genes in the body.This experiment mainly focuses on pax6 of Xenopus to VP of Bufo.raddei,the similarities and differences between the two kind of cells on cell proliferation,cycle regulation and the influence of cell fate determination,as well as the change of the related sites impact on the cell proliferation and its downstream,in order to predict which specific sites plays the main function.Phosphorylation sites prediction of PAX6 and VP protein and sequence alignment of them shows that the difference between them is in the structure of PST domain,and the phosphorylation sites or sites can be recognized on the phosphorylation of kinases reduces significantly in the VP.For its different phosphorylation site structure,we design point mutation plasmid and do the related experiment,we speculate that the 373 phosphorylation sites of the gene play the leading factor,the other phosphorylation site has the corresponding effect.