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件， 可以方便和快速使用的遗传程序正在被 开发 与这种方法相互交织的是DNA的特定主题， 转置正在研究，无论是为了基本了解 这是一个过程，并作为进一步基因应用的基础。 因为 E. coli中，这些研究开始于E.杆菌和 被进一步改造以应用于其他生物体，包括酵母 和高等真核细胞。 所使用的许多程序涉及特殊载体的构建， 转座子，可用于融合部分特征良好的基因， 其他基因的互补部分。 这种基因融合过程可以 用于鉴定基因片段或结构域， 可用于进一步的遗传和生物化学研究。 增加 基因融合的多功能性，正在寻找新类型的基因， 特性不同于最常用的基因，E.杆菌 β-半乳糖苷酶基因。 这些基因包括其他水解酶的基因 其像B-半乳糖苷酶一样具有宽范围的比色底物， 以及B-半乳糖苷酶基因和 其他类型的基因如氨基糖苷类耐药基因或胸苷类耐药基因 激酶。 转座因子Tn 3是转座因子研究和应用的热点 和噬菌体Mu转座子。 特别感兴趣的是 转座酶蛋白结合的转座子末端序列，以及 异常的转座结构，当这些末端 序列被改变了。 新的技术应用可以通过 转座子末端之间的各种遗传实体 例如启动子、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 结构有许多可能的应用，包括形成 各种类型的基因融合，DNA克隆完全在体内，定向DNA 克隆（步移），局部重组，定点DNA转移，新的 基因选择和快速DNA测序。 畸变研究 转座产品将增加我们的理解的过程， DNA转座，并可能允许开发新的应用程序， 转座因子 建议的其他实验包括测试 Tn 3和Mu元件在其他物种中的转座以扩展其用途 到更高的物种，因为已经与B-半乳糖苷酶基因融合 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.