Kroos L, Kuspa A, Kaiser D
Dev Biol. 1986 Sep;117(1):252-66. doi: 10.1016/0012-1606(86)90368-4.
Tn5 lac is a transposon that fuses the transcription of lacZ to exogenous promoters. We generated 2374 Tn5 lac insertion-containing strains of Myxococcus xanthus, a soil bacterium that undergoes multicellular development which culminates in the formation of spores. Thirty-six strains were identified that specifically increase beta-galactosidase expression at some particular time during development and these expression times range from minutes after starvation initiates development to 24 hr, when sporulation begins. Different maximum levels of beta-galactosidase expression were also observed and the maximum for many strains that begin beta-galactosidase expression late in development was observed only if spores were disrupted. Seven of the 36 strains display mild to severe defects in aggregation and/or sporulation, as did an additional five strains whose beta-galactosidase expression was not developmentally regulated. Restriction maps of the DNA adjacent to the Tn5 lac insertions that are developmentally regulated and/or cause developmental defects show that most of the 41 insertions are in different regions of the Myxococcus genome. The developmentally regulated Tn5 lac insertions described here provide a set of at least 29 new developmental markers for Myxococcus.
Tn5 lac是一种转座子,它将lacZ的转录与外源启动子融合。我们构建了2374株含有Tn5 lac插入片段的黄色粘球菌菌株,黄色粘球菌是一种土壤细菌,会经历多细胞发育,最终形成孢子。我们鉴定出36株菌株在发育的某些特定时间会特异性增加β-半乳糖苷酶的表达,这些表达时间从饥饿引发发育后的几分钟到孢子形成开始的24小时不等。还观察到β-半乳糖苷酶表达的不同最大水平,并且许多在发育后期开始β-半乳糖苷酶表达的菌株只有在孢子被破坏时才观察到最大值。36株菌株中有7株在聚集和/或孢子形成方面表现出轻度到重度的缺陷,另外5株β-半乳糖苷酶表达不受发育调控的菌株也是如此。与受发育调控和/或导致发育缺陷的Tn5 lac插入片段相邻的DNA的限制性图谱表明,41个插入片段中的大多数位于黄色粘球菌基因组的不同区域。这里描述的受发育调控的Tn5 lac插入片段为黄色粘球菌提供了一组至少29个新的发育标记。