Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces.
Natural and synthetic tetracycline-inducible promoters for use in the antibiotic-producing bacteria Streptomyces.
复制标题
天然和合成四环素诱导的启动子用于抗生素产生细菌链霉菌。
DOI：
10.1093/nar/gni086
复制
发表时间：
2005-05-25
影响因子：
14.9
通讯作者：
Smith, Margaret C M
中科院分区：
文献类型：
--
作者：
Rodriguez-Garcia, Antonio;Combes, Patricia;Perez-Redondo, Rosario;Smith, Matthew C A;Smith, Margaret C M
链霉菌属细菌是抗生素和其他药理活性化合物的主要生产者。这些细菌的遗传和生理操作对于新药发现和生产开发非常重要。任何“遗传工具包”的一个重要组成部分是可调节启动子的可用性。我们改编了转座子 Tn10 的四环素 (Tc) 阻遏物/操纵子 (TetR/tetO) 调节系统，用于链霉菌。合成的 Tc 可控启动子 (tcp) tcp830 在多种链霉菌属物种中都有活性，并且在添加 1–100 ng/ml 的脱水四环素 (aTc) 后观察到不同水平的诱导。天蓝色链霉菌含有一个由 TetR 同源物 (SCO0253) 调节的先天 Tc 可控启动子。天然和合成启动子在整个生长过程中都是活跃且可诱导的。使用表达荧光素酶的 luxAB 基因作为报告系统，我们表明 tcp830 可以获得高达 270 的诱导因子。确定了诱导剂对天蓝色链球菌生长的影响；以诱导最佳浓度（即 0.1–1 μg/ml）添加 aTc，与没有诱导剂的培养物相比，对生长速率没有影响，到滞后期略有增加。
Bacteria in the genus Streptomyces are major producers of antibiotics and other pharmacologically active compounds. Genetic and physiological manipulations of these bacteria are important for new drug discovery and production development. An essential part of any ‘genetic toolkit’ is the availability of regulatable promoters. We have adapted the tetracycline (Tc) repressor/operator (TetR/tetO) regulatable system from transposon Tn10 for use in Streptomyces. The synthetic Tc controllable promoter (tcp), tcp830, was active in a wide range of Streptomyces species, and varying levels of induction were observed after the addition of 1–100 ng/ml of anhydrotetracycline (aTc). Streptomyces coelicolor contained an innate Tc-controllable promoter regulated by a TetR homologue (SCO0253). Both natural and synthetic promoters were active and inducible throughout growth. Using the luxAB genes expressing luciferase as a reporter system, we showed that induction factors of up to 270 could be obtained for tcp830. The effect of inducers on the growth of S.coelicolor was determined; addition of aTc at concentrations where induction is optimal, i.e. 0.1–1 μg/ml, ranged from no effect on growth rate to a small increase in the lag period compared with cultures with no inducer.