可逆的蛋白磷酸化是细胞内信号传递的关键过程，几乎调控着细胞生理过程的各个方面。本项目基于南移养殖仿刺参（Apostichous japonicus Selenka）耐高温良种培育的实际需求，以仿刺参应答高温胁迫的内在分子响应机制为突破口，借助磷酸化蛋白富集和磷酸化蛋白组学等技术手段，发掘仿刺参应答高温胁迫的相关磷酸化修饰蛋白10个以上；随后，利用磷酸化Western blot检测差异磷酸化蛋白在高温胁迫前后的表达变化，并结合其在mRNA水平上的表达情况初步确认其与高温胁迫的联系；在此基础上，筛选1-2个关键磷酸化蛋白，采用酵母体外重组和GST pull-down策略，发掘其胞内的潜在结合蛋白2-10个。最后，借鉴高等生物相关蛋白磷酸化信号通路研究进展，勾勒出仿刺参应答高温胁迫的内在调控机制雏形。该研究不仅为刺参耐高温品系的选育提供新的思路和方向，同时为人们认识刺参夏眠的内在机制提供参照。

Reversible protein phosphorylation represents a key control mechanism in signal transmission and controls nearly all aspects of cellular physiology. In order to acheieve the goal to breed temperature-tolerance sea cucumber Apostichous japonicus strain in practice, the intrinsic response mechanism of sea cucumber against thermal stree was investigated focusing on phosphorylated proteins. Phosphoproteins enrichment techniques and phosphoproteomics were employed in the study and more than 10 temperature related phosphoproteins should be identified. These protein were further validated by subsequent phosphous Western blot and expression pattern analysis at mRNA level. After that, the GST fusion protein expression system was constructed and the recombinant proteins were produced in yeast cell DY150 for some vital members, and their potential binding proteins were further determined by GST pull-down assay and mass spectrometry. Combination with ongoing achievement on phosphoprotein signal pathway, phosphorylation-based signaling networks would be firstly delineated in this lower species. We hoped the present study would provide new insight into sea cucumber breeding process and unveil the mask for the species aestiviation from one aspect.