Abstract
Confocal laser scanning microscopy was used to observe the spatio-temporal expression of the pathway-specific gene redD during S. coelicolor cell cultivation. The corresponding mutant S. coelicolor lyqRY1522 carrying redD::eyfp in the chromosome was constructed. The temporal expression results of the fusion protein during submerged cultivation demonstrated that expression of redD began in the transition phase, continuing through the exponential growth phase to the stationary phase, and reached maximum in the stationary phase. On the other hand, redD was expressed only in substrate mycelia during solid-state culture, while aerial mycelia remained essentially non-fluorescent throughout culture. Results demonstrated that the expression pattern of redD coincides with that of the biosynthesis of the antibiotics during culture, revealing a direct correlation between the spatio-temporal distribution of regulatory gene expression and second metabolism.
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References
Beavis, A.J., Kalejta, R.F., 1999. Simultaneous analysis of the cyan, yellow and green fluoresent proteins by flow cytometry using single-laser excitation at 458 nm.Cytometry,37:68–73.
Bierman, M., Logan, R., O'Brien, K., Seno, E.T., Rao, R.N., Schoner, B.E., 1992. Plasmid cloning vectors for the conjugal transfer of DNA fromEscherichia coli toStreptomyces spp.Gene,146:43–49.
Brendan, P.C., Raphael, H.V., Stanley, F., 1996. FACS-optimized mutants of green fluorescence protein.Gene,173:33–38.
Fernandez-Moreno, M.A., Martinez, E., Caballero, J.L., Ichinose, K., Hopwood, D.A., Malpartida, F., 1994. DNA sequence and functions of theactVI region of the actinorhodin biosynthetic gene cluster ofStreptomyces coelicolor A3(2).Journal of Biological Chemistry,269(40):24854–24863.
Gary, M.D., Stephen, C.W., 1999. Cell-Cell Signaling in Bacteria. American Society for Microbiology Press, Washington D.C., p. 27–46.
Hopwood, D.A., Chater, K.F., Bibb, M.J., 1995. Genetics of antibiotic production inStreptomyces coelicolor A3(2), a model streptomyces.Biotechnology,28:65–102.
Huang, J., Lih, C.J., Pan, K.H., Cohen, S.N., 2001. Global analysis of growth phase responsive gene expression and regulation of antibiotic biosynthetic pathways in Streptomyces coelicolor using DNA microarrays.Genes & Development,15:3183–3192.
Kieser, T., Bibb, M.J., Chater, K.F., Hopwood, D.A., 2000. Practical Streptomyces Genetics. John Innes Foundation, Norwich, United Kingdom.
Kyung, Y.S., Hu, W.S., Sherman, D.H., 2001. Analysis of temporal and spatial expression of the CcaR regulatory element in the cephamycin C biosynthetic pathway using green fluorescent protein.Molecular Microbiology,40(3):530–541.
Prasher, D.C., Eckenrode, V.K., Ward, W.W., Prendergast, F.G., Cormier, M.J., 1992. Primary structure of theAequorea victoria green-fluorescent protein.Gene,111:229–233.
Sambrook, J., Fritsch, E.F., Maniatis, T., 1989. Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor Laboratory.
Takano, E., Gramajo, H.C., Strauch, E., Andres, N., White, J., Bibb, M.J., 1992. Transcriptional regulation of theredD transcriptional activator gene accounts for growth-phase-dependent production of the antibiotic undecylprodigiosin inStreptomyces coelicolor A3(2).Molecular Microbiology,6(19):2797–2804.
Yang, T.T., Sinai, P., Green, G., Kitts, P.A., Chen, Y.T., Lybarger, L., Chervenak, R., Patterson, G.H., Piston, D.W., Kain, S.R., 1998. Improved fluorescence and dual color detection with enhanced blue and green variants of the green fluorescent protein.Journal of Biological Chemistry,273:8212–8216.
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Project (No. 2004-527) supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, China
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Zhou, Lh., Li, Yq., Li, Yq. et al. Spatio-temporal expression of the pathway-specific regulatory gene redD in S. coelicolor. J Zheijang Univ Sci B 6, 464–469 (2005). https://doi.org/10.1631/jzus.2005.B0464
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DOI: https://doi.org/10.1631/jzus.2005.B0464