Abstract
A useful approach to the study of differentiation consists of isolating and characterizing the genes specifically expressed in the various stages and then studying their regulation. This chapter describes the methodology used in our laboratory to clone sequences from stage-specific mRNAs of Physarum polycephalum. It involved the preparation of intact poly (A)+ RNA and the construction of cDNA libraries for four developmental stages (amoebae, plasmodia, spores, and spherules). These libraries were then screened by differential hybridization, and the specificity of the positive clones was confirmed by Northern blot hybridization. The quality of the results obtained at each step was controlled, and the methodology was found to be of general use in the identification of Physarum stage-specific mRNAs.
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© 1986 Plenum Press, New York
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Pallotta, D., Bernier, F., Hamelin, M., Martel, R., Lemieux, G. (1986). cDNA Cloning Of Physarum polycephalum Stage-Specific mRNAs. In: Dove, W.F., Dee, J., Hatano, S., Haugli, F.B., Wohlfarth-Bottermann, KE. (eds) The Molecular Biology of Physarum polycephalum . NATO ASI Series, vol 106. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2203-0_24
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DOI: https://doi.org/10.1007/978-1-4613-2203-0_24
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