Hahn D R, Solenberg P J, McHenney M A, Baltz R H
Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285.
J Ind Microbiol. 1991 Jun;7(4):229-34. doi: 10.1007/BF01577649.
To expand the application of molecular genetics to many different streptomycete species, we have been developing two potentially widely applicable methodologies: transposon mutagenesis and plasmid transduction. We constructed three transposons from the Streptomyces lividans insertion sequence IS493. Tn5096 and Tn5097 contain an apramycin resistance gene inserted in different orientations between the two open reading frames of IS493. These transposons transpose from different plasmids into many different sites in the Streptomyces griseofuscus chromosome and into its resident linear plasmids. Tn5099 contains a promoterless xylE gene and a hygromycin-resistance gene inserted in IS493 close to one end. Tn5099 transposes in S. griseofuscus giving operon fusions in some cases that drive expression of the xylE gene product, catechol deoxygenase, giving yellow colonies in the presence of catechol. We have also developed plasmid vectors that can be transduced into many streptomycete species by bacteriophage FP43. We describe the characterization of FP43 and mapping of several bacteriophage functions. The region of cloned FP43 DNA essential for plasmid transduction includes the origin for headful packaging.
为了将分子遗传学应用扩展到许多不同的链霉菌物种，我们一直在开发两种可能广泛适用的方法：转座子诱变和质粒转导。我们从淡紫灰链霉菌插入序列IS493构建了三种转座子。Tn5096和Tn5097含有一个安普霉素抗性基因，该基因以不同方向插入IS493的两个开放阅读框之间。这些转座子从不同质粒转座到灰褐链霉菌染色体的许多不同位点及其常驻线性质粒中。Tn5099含有一个无启动子的xylE基因和一个潮霉素抗性基因，它们插入到靠近IS493一端的位置。Tn5099在灰褐链霉菌中转座，在某些情况下产生操纵子融合，驱动xylE基因产物儿茶酚脱氧酶的表达，在儿茶酚存在下产生黄色菌落。我们还开发了可以通过噬菌体FP43转导到许多链霉菌物种中的质粒载体。我们描述了FP43的特性以及几种噬菌体功能的定位。质粒转导所必需的克隆FP43 DNA区域包括头部包装的起始位点。
