GENE FUSION AND THE STUDY OF DNA TRANSPOSITION

为了研究许多不同类型的基因和遗传元素， 可以方便快捷地使用的基因程序正在被 发展起来的。与这种方法交织在一起的是DNA的特定主题 人们正在研究移位，这既是为了从根本上理解 并将其作为进一步基因应用的基础。因为 在大肠杆菌中操作的简易性，这些研究是从大肠杆菌和 被进一步改造用于包括酵母在内的其他生物体 和高等真核细胞。 所使用的许多过程涉及特殊向量的构造和 转座子可以用来融合特征良好的基因的部分 其他基因的互补部分。这种基因融合过程可以是 用于识别基因片段或结构域，以及由此产生的基因融合 可用于进一步的遗传和生化研究。为了增加 基因融合的多功能性，人们正在寻找新类型的基因 特性不同于最常用的基因--大肠杆菌 β-半乳糖苷酶基因。这些包括其他水解酶的基因。 它们和B-半乳糖苷酶一样，具有广泛的比色底物，如 以及B-半乳糖苷酶基因和 其他类型的基因，如氨基糖苷耐药性或胸腺嘧啶核苷 激活剂。 转座元件的研究和应用主要集中在Tn3 以及噬菌体Mu转座子。特别值得关注的是 转座酶蛋白结合的转座子末端序列，以及 当这些末端形成异常转位结构时 序列被改变。新的技术应用程序可以通过 通过在转座子末端之间整合不同的遗传实体来实现 如启动子、结构基因、复制子、重组位点、 包装地点和转移来源。由此产生的转座 构造有许多可能的应用，包括形成 各种类型的基因融合，完全在体内克隆DNA，定向DNA 克隆(行走)、局部重组、定点DNA转移、新 基因选择的类型，以及快速DNA测序。反常现象的研究 移位产品将增加我们对 DNA转座，并可能允许开发新的应用 转座因子。建议进行的其他实验包括测试 Tn3和Mu元素在其他物种中的转位以扩展其用途 对于高等物种，就像对B-半乳糖苷酶基因融合所做的那样 技术

In order to study many different types of genes and genetic elements, genetic procedures that can be used conveniently and rapidly are being developed. Intermeshed with this approach the specific subject of DNA transposition is being studied, both for a fundamental understanding of the process and as a basis for further genetic applications. Because of the ease of manipulations in E. coli, these studies are started in E. coli and are further engineered for applications in other organisms including yeast and higher eukaryotic cells. Many of the procedures used involve the construction of special vectors and transposons that can be used to fuse parts of well-characterized genes to the complementary parts of other genes. This gene fusion process can be used to identify gene segments or domains, and the resulting gene fusions can be used for further genetic and biochemical studies. To increase the versatility of gene fusions, new types of genes are being sought with properties different from the most commonly used gene, the E. coli B-galactosidase gene. These include genes for other hydrolytic enzymes which like B-galactosidase have a wide range of colorimetric substrates, as well as composite or hybrid genes between the B-galactosidase gene and other types of genes such as for aminoglycoside resistance or thymidine kinase. Studies and applications of transposable elements are focusing on the Tn3 and the bacteriophage Mu transposons. Of particular interest are the transposon terminal sequences where the transposase proteins bind, and the aberrant transposition structures that are formed when these terminal sequences are altered. New technical applications can be developed with transposon termini by incorporating between them various genetic entities such as promoters, structural genes, replicons, recombination sites, packaging sites, and transfer origins. The resulting transposable constructs have many possible applications, including the formation of various types of gene fusions, DNA cloning entirely in vivo, directed DNA cloning (walking), localized recombination, site directed DNA transfer, new types of genetic selections, and rapid DNA sequencing. Studies of aberrant transposition products will increase our understanding of the process of DNA transposition and may allow the development of new applications for transposable elements. Additional experiments proposed involve testing transposition of Tn3 and Mu elements in other species to extend their uses to higher species, as has been done with the B-galactosidase gene-fusion technology.