Cermák L, Novotná J, Ságova-Marecková M, Kopecký J, Najmanová L, Janata J
Institute of Microbiology, 142 20 Prague, Czechia.
Folia Microbiol (Praha). 2007;52(5):457-62. doi: 10.1007/BF02932104.
The first insight into celesticetin biosynthetic gene cluster of S. caelestis is presented. The genomic DNA of producing strain was digested, digoxigenin-labeled and hybridized with a set of probes designed according to S. lincolnensis gene sequences. Genes with high homology to the lincomycin biosynthetic genes coding for the predicted common parts of the pathway were identified in S. caelestis. Then, genomic DNA of S. caelestis treated by a multiple digestion was hybridized with five digoxigenin-labeled probes to construct a rough restriction map. Two consecutive islands formed by the genes with a putative function in biosynthesis of the shared saccharide moiety revealed an organization similar to the lincomycin biosynthetic gene cluster. The celesticetin cluster was mapped and essential information was obtained for subsequent steps, i.e. isolation and sequence analysis of the cluster.
首次报道了对天蓝链霉菌中天蓝菌素生物合成基因簇的研究。将产生菌株的基因组DNA进行酶切、地高辛标记,并与根据林肯链霉菌基因序列设计的一组探针杂交。在天蓝链霉菌中鉴定出与编码该途径预测共同部分的林可霉素生物合成基因具有高度同源性的基因。然后,将经多次酶切处理的天蓝链霉菌基因组DNA与五个地高辛标记的探针杂交,构建粗略的限制性图谱。由在共享糖部分生物合成中具有推定功能的基因形成的两个连续区域显示出与林可霉素生物合成基因簇相似的组织形式。绘制了天蓝菌素基因簇图谱,并获得了后续步骤(即基因簇的分离和序列分析)所需的重要信息。