caa a22 4500 605499276 CHVBK 20210128100550.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6255-9 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6255-9 Capturing the target genes of BldD in Saccharopolyspora erythraea using improved genomic SELEX method [Elektronische Daten] [Hang Wu, Yongrong Mao, Meng Chen, Hui Pan, Xunduan Huang, Min Ren, Hao Wu, Jiali Li, Zhongdong Xu, Hualing Yuan, Ming Geng, David Weaver, Lixin Zhang, Buchang Zhang] BldD (SACE_2077), a key developmental regulator in actinomycetes, is the first identified transcriptional factor in Saccharopolyspora erythraea positively regulating erythromycin production and morphological differentiation. Although the BldD of S. erythraea binds to the promoters of erythromycin biosynthetic genes, the interaction affinities are relatively low, implying the existence of its other target genes in S. erythraea. Through the genomic systematic evolution of ligands by exponential enrichment (SELEX) method that we herein improved, four DNA sequences of S. erythraea A226, corresponding to the promoter regions of SACE_0306 (beta-galactosidase), SACE_0811 (50S ribosomal protein L25), SACE_3410 (fumarylacetoacetate hydrolase), and SACE_6014 (aldehyde dehydrogenase), were captured with all three BldD concentrations of 0.5, 1, and 2μM, while the previously identified intergenic regions of eryBIV-eryAI and ermE-eryCI plus the promoter region of SACE_7115, the amfC homolog for aerial mycelium formation, could be captured only when the BldD's concentration reached 2μM. Electrophoretic mobility shift assay (EMSA) analysis indicated that BldD specifically bound to above seven DNA sequences, and quantitative real-time PCR (qRT-PCR) assay showed that the transcriptional levels of the abovementioned target genes decreased when bldD was disrupted in A226. Furthermore, SACE_7115 and SACE_0306 in A226 were individually inactivated, showing that SACE_7115 was predominantly involved in aerial mycelium formation, while SACE_0306 mainly controlled erythromycin production. This study provides valuable information for better understanding of the pleiotropic regulator BldD in S. erythraea, and the improved method may be useful for uncovering regulatory networks of other transcriptional factors. Springer-Verlag Berlin Heidelberg, 2014 Saccharopolyspora erythraea nationallicence Improved genomic SELEX method nationallicence BldD nationallicence amfC nationallicence SACE_0306 nationallicence Wu Hang Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Mao Yongrong Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Chen Meng Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Pan Hui Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Huang Xunduan Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Ren Min Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Wu Hao Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Li Jiali Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Xu Zhongdong School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut Yuan Hualing School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut Geng Ming School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut Weaver David Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Zhang Lixin Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Zhang Buchang Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2683-2692 0175-7598 99:6<2683 2015 99 253 https://doi.org/10.1007/s00253-014-6255-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6255-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wu Hang Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Mao Yongrong Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Chen Meng Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Pan Hui Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Huang Xunduan Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Ren Min Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Wu Hao Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Li Jiali Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Xu Zhongdong School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut NATIONALLICENCE 700 1- Yuan Hualing School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut NATIONALLICENCE 700 1- Geng Ming School of Life Sciences, Hefei Normal University, 234000, Hefei, China aut NATIONALLICENCE 700 1- Weaver David Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Zhang Lixin Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 700 1- Zhang Buchang Institute of Health Sciences, School of Life Sciences, Anhui University, 230601, Hefei, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/6(2015-03-01), 2683-2692 0175-7598 99:6<2683 2015 99 253