INVESTIGATION OF MU TRANSPOSITION IN VITRO

MU是一种细菌病毒，它利用基因转座整合和 复制它的DNA。Mu是已知的最活跃的转座元素， 在裂解循环中，Mu使用大肠杆菌复制机器来 转座到细菌染色体上的100个不同的位置 小时。转座所需的病毒成分包括DNA序列 在病毒的两端以及Mu基因A和B的产物上，我们的龙 Range目标是剖析小鼠的生化机制 换位。我们最近开发了一种体外系统来进行 高效的Mu转位-复制。我们计划在体外使用这种技术 转座中间体的表征和Mu的提纯系统 转座蛋白。在这些研究过程中，我们将确定 Mu复制的起源(S)，定义了DNA的启动机制 在原产地合成，并测定其生化性质 进行转座的蛋白质。换位的过程现在是 在耐药基因和质粒的传播中被认识到 细菌，在原核和真核染色体的DNA重排中， 在致癌病毒的发育周期中。然而， 基因易位的生物学影响才刚刚开始 感激不尽。这些研究将极大地提高我们对 参与这一现象的蛋白质和遗传序列的功能。

Mu is a bacterial virus that uses genetic transposition to integrate and replicate its DNA. Mu is the most active transposable element known and during the lytic cycle Mu uses the E. coli replication machinery to transpose to 100 different sites in the bacterial chromosome within an hour. The viral elements necessary for transposition include DNA sequences at both ends of the virus and the products of Mu genes A and B. Our long range objective is to dissect the biochemical mechanism of Mu transposition. We recently developed an in vitro system for carrying out efficient Mu transposition-replication. We plan to use this in vitro system to characterize transposition intermediates and to purify Mu transposition proteins. In the course of these studies we will identify the origin(s) of Mu replication, define the mechanism of initiation of DNA synthesis at the origin, and determine the biochemical properties of the proteins that carry out transposition. The process of transposition is now recognized in the dissemination of drug resistance genes and plasmids in bacteria, in DNA rearrangements of prokaryotic and eukaryotic chromosomes, and in the developmental cycle of oncogenic viruses. However, the biological impact of genetic transposition is only beginning to be appreciated. These studies will significantly advance our knowledge of the functions of the proteins and genetic sequences involved in this phenomenon.