METHODOLOGY DEVELOPMENT FOR THE ELUCIDATION OF PHOSPHORYLATION SITES ON PROTEIN

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。我们设计了一个用于将酵母中内源蛋白启动子同源替换为诱导型启动子的PCR模板。表达了来自这些可诱导启动子的内源性激酶的几种蛋白质底物，并对蛋白质上的磷酸化位点进行了定位。我们正在通过定位这些蛋白质中的几个磷酸化位点来测试这项技术，这些蛋白质包括MCM3，Bud3，boi2，lte1，ned1。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. we designed a PCR template for homologous replacement of the endogenous protein promoters in yeast to inducible promoters. Expressed several protein substrates of endogenous kinases from these inducible promoters and in order to map phosphorylation sites on the proteins. We are testing the technique by mapping phosphorylation sites on several of these proteins including mcm3, bud3, boi2, lte1, ned1.