ILLEGITIMATE RECOMBINATION BY DRUG RESISTANCE ELEMENTS

转座因子在介导DNA重排中起主要作用 在原核生物内部和之间。 我们建议调查 原核插入转座的机制与调控 序列的 我们的分析将特别强调转座子Tn 10和 它的组成部分IS 10序列，但也将分析IS 26的某些方面 所以是5。 现提出以下七个具体项目： (1)IS 10体外转座的生化分析。 (2)IS 10转座酶蛋白的分离及遗传和物理分析 和转座酶突变体的一般和特定类别的表征。 (3)IS 26和IS 5转座的遗传和生化比较， 这就是IS 10。 (4)转座子供体分子的命运的遗传分析， 换位 (5)IS 10插入特异性的剖析及其作用的探讨 IS 10和IS 26的区域特异性DNA熔解。 (6)IS 10反义RNA介导的翻译调控机制 RNA-OUT和RNA-IN之间的配对和RNA-IN的结合的体外分析 核糖体到配对和未配对的RNA。 (7)dam甲基化对IS 10转座的调节：转座动力学 在体内和体外对非复制和复制DNA的甲基化。

Transposable elements play a major role in mediating DNA rearrangements within and between prokaryotic organisms. We propose to investigate the mechanism and regulation of transposition by prokaryotic insertion sequences. Our analysis will particularly emphasize transposon Tn10 and its component IS10 sequences, but will also analyze some aspects of IS26 and IS5. Seven specific projects are proposed below: (1) Biochemical analysis of IS10 transposition in vitro. (2) Genetic and physical analysis of IS10 transposase protein: isolation and characterization of general and specific classes of transposase mutants. (3) Genetic and biochemical comparison of transposition by IS26 and IS5 to that of IS10. (4) Genetic analysis of the fate of the transposon donor molecule during transposition. (5) Dissection of IS10 insertion specificity and investigation of the role of DNA melting in regional specificity of IS10 and IS26. (6) Mechanism of IS10-mediated translational control by anti-sense RNA: in vitro analysis of pairing between RNA-OUT and RNA-IN and binding of ribosomes to paired and unpaired RNAs. (7) Regulation of IS10 transposition by dam methylation: kinetics of methylation on non-replicating and replicating DNA in vivo and in vitro.