CORE--PEPTIDE SYNTHESIS AND ANALYSIS

该共享设施的广泛目标是使用Edman化学和肽合成服务进行肽测序，以及氨基酸组成分析，随时可供癌症中心的研究人员使用。分离的蛋白质的鉴定，无论是以前描述的还是以前未报道的蛋白质，都是许多研究项目的重要组成部分，通常最好通过N-末端测序来实现。用于分子生物学研究的寡核苷酸探针的设计通常基于从N-末端序列分析获得的有限肽序列。合成肽有许多需要：经常与蛋白质结合并用作特异性免疫原。癌症中心的肽分析和合成共享设施每年对150-250种肽进行测序，并合成90-120种肽。其中70%以上是为癌症中心的教师提供的。该设施目前的使用程度表明了对这些服务的需要和价值。他们在帮助实现许多癌症中心研究项目的目标方面的重要性在本更新提案的其他地方进行了概述。最近，已经建立了用于获得来自电印迹到PVDF的蛋白质的肽的内部N-末端序列的方法，并在该设施中提供。此外，该基金的网页也得到了加强，现在包括了所提供服务和所采用方法的全面说明。该机制使用的方法已经成熟，不再会迅速改变。与质谱共享设施不断互动，以确保在每种实验情况下推荐最合适和最有效的技术。此外，通常的做法是利用MALDI TOF MS分析来验证合成的肽的组成，并选择最理想的肽进行序列分析。可以预见，MS将越来越多地用于许多测序任务。

The broad objective of this Shared Facility is to make peptide sequencing using Edman chemistry and peptide synthesis services, as well as amino acid composition analysis, readily available to Cancer Center researchers. Identification of an isolated protein, whether a previously described or previously unreported protein, is an essential part of many research projects and commonly is best achieved by N-terminal sequencing. The design of oligonucleotide probes for molecular biological studies is most often based upon limited peptide sequence obtained from N-terminal sequence analysis. There are many needs for synthesis of peptides: frequently to be conjugated to protein and used as specific immunogens. The Cancer Center's Shared Facility for Peptide Analysis & Synthesis sequences 150-250 peptides and synthesizes 90-120 peptides annually. More than 70% of these services are for Cancer Center faculty. The present extent of usage of the Facility is indicative of the need for and value of these services. Their importance in helping to achieve the goals of many Cancer Center research projects is outlined elsewhere in this renewal proposal. Recently, methodologies for obtaining internal N-terminal sequence of peptides derived from proteins electroblotted to PVDF have been established and made available in this Facility. Also, the Facility' web page has been enhanced and now includes full descriptions of services offered and methodologies to be employed. The methodologies used by the Facility have matured and are no longer subject to rapid change. There is constant interaction with the Mass Spectrometry Shared Facility to ensure that the most appropriate and efficient techniques are recommended in each experimental situation. In addition, it is usual practice to utilize MALDI TOF MS analysis to verify the composition of peptides synthesized and to select the most desirable peptides for sequence analysis. It is predictable that MS will be used increasingly for many sequencing tasks.