ANALYSIS AND SORTING OF BACTERIAL LIBRARY EXPRESSING GFP

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 GFP已被用作融合蛋白折叠和溶解的报告分子。以GFP为支架构建的文库由折叠良好的蛋白质和未折叠的蛋白质组成。折叠不好的GFP在选择过程中往往会给出背景。因此，我们希望用流式细胞术根据亮度将文库(108-109)分成三组。将从文库中选择结核或其他蛋白质的结合体。特定粘结剂的激发光谱和发射光谱将通过荧光法(Spex，Fluorolog)测定。在对粘合剂进行表征之后，将使用FacsCalibur测试它们在真核细胞上的有效性和分布。

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. GFP has been used as reporter for folding and solubility of a fusion protein. Libraries made using GFP as scaffold consists of well folded as well as unfolded protein. GFP that is not folded well tends to give background during the selection. Therefore, we wish to sorted library (108-109) into three different groups based on the brightness using flow cytometry. Binders to TB or other proteins will be selected from the library. Excitation and emission spectra of specific binders will be determined by fluorimetry (Spex, Fluorolog). Following characterization of binders, their efficacy and distribution on/in eukoryotic cells will be tested using the FacsCalibur.