FLUORESCENT ACTIVATED CELL SORTING FOR ENZYME SCREENING

描述：（改编自申请人的摘要）该项目的第一阶段是 旨在开发一种基于 在荧光激活细胞分选的发现独特的，强大的， 特定的酶催化剂。这些酶对于那些 用于生产关键化合物的手性二氢吡喃酮的成本有效的合成 药品、生物中间体和天然产物以及用于 基于酶的组合药物筛选。流式细胞仪筛查系统开发 在第一阶段，每秒处理25，000个克隆， 多种酶类型并实现显著的发现率。 在第一阶段的基础上，第二阶段的工作将致力于完善和扩大 流式细胞仪系统的筛选能力，包括几个额外的 基于生物催化剂的工业需要的酶的种类。此外 将改进切除方案以访问环境基因组文库 由大于1 x 10的8次方成员和单细胞PCR组成 将被改进以捕获编码有毒产物如蛋白酶的DNA。 拟定商业应用：不可用

DESCRIPTION: (adapted from applicant's abstract) Phase I of this project was aimed at the development of an extremely high throughout screening system based on Fluorescence activated cell sorting for the discovery of unique, robust, and specific enzyme catalysts. These enzymes are especially valuable for the cost-effective synthesis of chiral synthons for use in production of key pharmaceuticals, biological intermediates and natural product as well as for enzyme based combinatorial drug screening. The FACS screening system developed in Phase I handles 25,000 clones per second, incorporates assay methods for multiple enzyme types and realizes significant discovery rates. Building upon Phase I, Phase II efforts will be directed to refine and expand the screening capabilities for the FACS system to include several additional classes of enzymes based upon industrial needs for biocatalysts. In addition excision protocols will be improved to access environmental genomic libraries consisting of greater than 1 x 10 to the 8th power members and single cell PCR will be refined to capture DNA that encodes toxic products such as proteases. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE