NOVEL SCREEN FOR RESTRICTION ENDONUCLEASES

限制性内切酶是基因工程的关键试剂。 虽然已经有两百多个具体的东西，但有 是对识别新序列的更多的需求。例如, 识别八核苷酸序列的其他酶将是 对作图很有用，许多疾病基因包含突变 如果适当的限制酶是可以很容易检测到的 可用。目前的生化筛查方法效率低下， 很费力，但很少成功。这些化验发现了局限性 仅在所检测的四种微生物中的一种中的酶和 发现新的特异性显著下降(只有3在 去年)。目前的DNA测序工作表明，所有 细菌至少有一个，可能还有更多的限制- 修改(RM)系统。这项提议提供了一种新的， 基于计算的方法，筛选每日更新的 潜在限制酶基因的GenBank，并有助于 找出可能有新的特殊性的那些。候选人将是 进行了生化筛选以验证预测。新的特性 将被充分描述并提供出售。 建议的商业应用这项研究将导致 新型限制性内切酶的发现 具体细节。它还将识别一些克隆和序列 以前未克隆的酶。新的酶将出售给这两家公司 向生物技术行业和学术研究界致敬。

Restriction endonucleases are key reagents for genetic engineering. Although more than two hundred specificities already exist, there is a need for more that recognize novel sequences. For instance, additional enzymes recognizing octanucleotide sequences would be useful for mapping and many disease genes contain mutations that could easily be detected if an appropriate restriction enzyme were available. Current biochemical screening methOds are inefficient, laborious and rarely successful. These assays find restriction enzymes in only one of four microorganisms examined and the rate of finding new specificities has dropped dramatically (only 3 in the last year). Current DNA sequencing efforts suggest that all bacteria possess at least one, and probably many more, Restriction- Modification (RM) systems. This proposal offers a new, computationally-based approach that screens the daily updates of GenBank for potential restriction enzyme genes and helps to identify those that may have new specificities. Candidates will be screened biochemically to test the predictions. New specificities will be fully characterized and offered for sale. PROPOSED COMMERCIAL APPLICATIONS This research will lead to the discovery of new restriction endonucleases with novel specificities. It will also identify clones and sequences for some previously uncloned enzymes. The new enzymes will be sold both to the Biotechnology industry and to the academic research community.