ANALYSIS AND SORTING OF BACTERIAL LIBRARY EXPRESSING GFP

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 GFP已被用作融合蛋白的折叠和溶解性的报告基因。使用GFP作为支架制备的文库由良好折叠以及未折叠的蛋白质组成。没有很好折叠的GFP倾向于在选择期间提供背景。因此，我们希望使用流式细胞术基于亮度将文库（108-109）分选成三个不同的组。 将从文库中选择TB或其他蛋白质的结合剂。将通过荧光测定法（Spex，Fluorolog）测定特定结合剂的激发和发射光谱。在表征结合剂后，将使用FacsCalibur检测其在真核细胞上/中的功效和分布。

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.