PROTEIN/PEPTIDE SEQUENCER AND SYNTHESIZER

我们请求购买一个集成的微型测序系统(型号 PI 2090E，波顿仪器公司)和多肽合成器(9050型， 米利根/生物搜索)。这些工具的选择是经过仔细考虑的 自1989年3月我们首次应用以来，进行了研究和重新评估。 我们相信，根据现有的数据，他们将是最好的 满足我们四名结缔组织研究人员的用户群的需求。 主要和次要的大部分多肽测序和合成任务 用户同样关心连接词的结构和交互作用的问题 组织元素(如蛋白多糖和胶原蛋白)。其中一个特殊的 UAB的研究优势是在连接领域。尽管具有关联性 纸巾工作者与不同的部门结盟，许多人需要并将 从针对他们的问题的肽/测序设施中受益。我们 预计需要80%的定序器操作时间 用于主要用户的项目。展示的许多多肽将是 通过电印迹将其固定到PVDF膜上。这个 测序仪适用于对这种形式的样品进行排序，并根据需要 敏感性(例如5微克蛋白质，相对分子质量50 KDa，为0.1 摩尔/周期)。有必要对测序方案进行更改，以 确定糖基化位点，这是我们经常需要做的。这个 选定的仪器具有允许这一点的必要灵活性。我们 预计主要用户将需要大约75%的运行时间 合成器。我们决定使用FMOC化学，它有几个 优势。例如，不需要像HF这样的试剂。和 已经开发出合成的多肽可以偶联到 系列片。在我们看来，所选择的多肽合成器是最 市场上用途广泛、经济实惠的工具。

We are requesting purchase of an Integrated Micro-sequencing System (Model PI 2090E, Porton Instruments, Inc.) and a Peptide Synthesizer (Model 9050, Milligen/Biosearch). The choice of these instruments has been carefully researched and reappraised since our original application of March, 1989. We are convinced, based upon data presently available, that they will best serve the needs of our user group of four connective tissue researchers. Most of the peptide sequencing and synthesis tasks for major and minor users alike relate to questions of structure and interactions of connective tissue elements (eg. proteoglycans and collagens). One of the particular research strengths of UAB is in the connective field. Although connective tissue workers are allied with different departments, many need and will benefit from a peptide/sequencing facility targeted to their problems. We predict that 80% of the operating time of the sequencer will be required for the major users' projects. Many of the peptides presented will be ready-immobilized through electro-blotting to PVDF membranes. The sequencer is suited to sequencing samples in this form and at the required sensitivity (eg. 5 microgram of protein, MW 50 KDa, which is 0.1 nmole/cycle). Changes in the sequencing protocol will be necessary to identify glycosylation sites, which we'll frequently need to do. The selected instrument has the necessary flexibility to permit this. We expect the major users to require about 75% of the operating time of the synthesizer. We have decided to use FMOC chemistry, which has several advantages. For example, there is no need for reagents like HF. And techniques have been developed where synthesized peptides can be coupled in series. The peptide synthesizer selected is, in our opinion, the most versatile and economic instrument on the market.