U.S.-Federal Republic of Germany Cooperative Research: The Influence of Phosphorothioate on the Action of Restriction Endonucleases

该奖项支持贡扎加的凯·纳卡马耶博士的研究访问 马克斯·普朗克实验医学研究所Max Planck Institute for Experimental Medicine Goettingen，FRG，以获得核苷酸合成的经验 并与弗里茨·埃克斯坦博士合作，他是该领域的领导者， 他是核酸化学的先驱， 硫代磷酸酯。 他们的合作研究将集中在使用 硫代磷酸酯在重组DNA的改进程序中的应用 操纵;具体地说，他们感兴趣的是 通过限制性酶切掺入DNA的硫代磷酸酯 核酸内切酶。 在之前的合作中，他们开发了一种简单的 在分离的细胞中产生位点特异性突变的有效方法 DNA片段或基因。 然而， 技术是需要一个特定的DNA序列的限制 在一些实施方案中，所述核酸内切酶优选在预期的核酸内切酶的位点附近使用。 突变他们现在计划研究IIS类限制性内切酶 作为指导突变和基因定位的策略， 通过内切核酸酶产生切口。 这将使它们的突变 该技术几乎适用于任何DNA和DNA序列。 诱变技术的发展已成为必不可少的， 蛋白质结构-功能等领域的研究 关系、基因控制和调节以及DNA/RNA结构。 基因分离、表征和操作是基础 许多生物技术的组成部分。 任何进一步的技术， 在基础研究和应用研究方面，这项工作将继续具有重大价值。

This award supports a research visit by Dr. Kay Nakamaye of Gonzaga University to the Max Planck Institute for Experimental Medicine in Goettingen, FRG, to gain experience in the synthesis of nucleotides and to collaborate with Dr. Fritz Eckstein, a leader in the field of nucleic acid chemistry who has pioneered the use of phosphorothioates. Their cooperative research will focus on the use of phosphorothioates in improved procedures for recombinant DNA manipulation; specifically, they are interested in the effect of phosphorothioates incorporated into DNA on cleavage by restriction endonucleases. In previous collaboration, they developed a simple yet efficient method of producing site-specific mutations in isolated DNA fragments or genes. However, one of the limitations of the technique is the need for a specific DNA sequence for the restriction endonuclease to be used, preferably near the site of the intended mutation. They now plan to study class IIS restriction endonucleases as a strategy for directing both the mutation and the position of nicking by the endonuclease. This would make their mutagenesis technique nearly universal for any DNA and DNA sequence. Development of mutagenesis techniques has become essential to investigations in areas such as protein structure-function relationships, gene control and regulation, and DNA/RNA structure. Gene isolation, characterization and manipulation are fundamental components of many biotechnologies. Any further techniques for such work will continue to have great value in basic and applied research.