Summary
Broad-bean (Vicia faba) chloroplast DNA (cpDNA) was isolated and characterized. The intact DNA is circular and has a molecular weight of 79.8x 106 dalton. Electron microscopic analysis of self-annealed intact single-strand circles show that it does not have a large double-stranded inverse repeat as seen in spinach chloroplast DNA. Only one ribosomal RNA gene (one set of 16S and 23S rRNA sequences) was found in preparations of R-loops between the Vicia rRNA and cpDNA circles. A restriction enzyme map for SalI and KpnI was derived by comparing the partial denaturation pattern of the fragments with the pattern of the intact circle. The map was confirmed by gel analysis. The ribosomal RNA gene was localized on the SalI fragment 3b by R-loop analysis. SalI fragment 1a although it contains a G-C rich region did not form R-loops with rRNA. Partial denaturation patterns of spinach cpDNA circles and BglI fragments were determined and from this the position of the fragments mapped. This confirmed the reliability of these methods for the arrangement of restriction enzyme fragments along circular molecules. The structures of the two cpDNAs were compared.
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References
Atchison, B.A., Whitfeld, P.R., Bottomley, W.: Comparison of chloroplast DNAs by specific fragmentation with EcoRI endonuclease. Mol. Gen. Genet. 148, 263–269 (1976)
Bedbrook, J.R., Bogorad, L.: Endonuclease recognition sites mapped on Zea mays chloroplast DNA. Proc. Natl. Acad. Sci. U.S.A. 73, 4309–4313 (1976)
Bedbrook, J.R., Kolodner, R.: The structure of chloroplast DNA. Annual Review of Plant Physiology, vol. 30, pp. 593–620 (1979)
Bedbrook, J.R., Kolodner, R., Bogorad, L.: Zea mays chloroplast ribosomal RNA genes are part of a 22,000 base pair inverted repeat. Cell 11, 739–749 (1977)
Chow, L.T., Roberts, J.M., Lewis, B., Broker, T.R.: A map of cytoplasmic RNA transcripts from lytic adenovirus type 2 determined by electron microscopy of RNA:DNA hybrids. Cell 11, 819–836 (1977)
Crouse, E.J., Schmitt, J.M., Bohnert, H.-J., Gordon, K., Driesel, A.J., Herrmann, R.G.: Intramolecular compositional heterogeneity of Spinacia and Euglena chloroplast DNAs. In: Chloroplast development (G. Akoyunoglou, ed.) pp. 565–572. Amsterdam: Elsevier/North-Holland Biomedical Press 1978
Davis, R.W., Simon, M., Davidson, N.: Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. In: Methods of enzymology (L. Grossmann and K. Moldave, eds.) vol. 21, pp. 413–428. New York: Academic Press 1971
Gray, P.W., Hallick, R.B.: Physical mapping of Euglena gracilis chloroplast DNA and ribosomal RNA gene region. Biochem. 17, 284–289 (1978)
Jenni, B., Stutz, E.: Analysis of Euglena gracilis chloroplast DNA. FEBS Lett. 102, 95–99 (1979)
Koller, B., Delius, H., Buenemann, H., Mueller, W.: The isolation of DNA from agarose gels by electrophoretic elution onto malachite green- polyacrylamide columns. Gene 4, 227–239 (1978)
Kolodner, R., Tewari, K.K.: The molecular size and conformation of the chloroplast DNA from higher plants. Biochim. Biophys. Acta 402, 372–390 (1975)
Kolodner, R., Tewari, K.K.: Denaturation mapping studies on the circular chloroplast deoxyribonucleic acid from pea leaves. J. Biol. Chem. 250, 4888–4895 (1975)
Kolodner, R., Tewari, K.K.: Physical studies on the size of the covalently closed circular chloroplast DNA from higher plants. Biochim. Biophys. Acta 447, 144–155 (1976)
Kolodner, R., Tewari, K.K.: Inverted repeats in chloroplast DNA from higher plants. Proc. Natl. Acad. Sci. U.S.A. 76, 41–45 (1979)
Mulder, C., Arrand, J.R., Delius, H., Keller, W., Petterson, U., Roberts, R.J., Sharp, P.A.: Cleavage maps of DNA from adenovirus types 2 and 5 by restriction endonucleases EcoRI and HpaI. Cold Spring Harbor Symp. Quant. Biol. 39, 397–400 (1975)
Pascoe, M.J., Ingle, J.: Distinction between nuclear satellite and chloroplast DNA in higher plants. Plant Physiol. 62, 975–977 (1978)
Portmann, R., Koller, Th.: The divalent cation method for protein-free spreading of nucleic acid molecules. Sixth European Congr. Electron Microscopy Jerusalem, 2, 546–548. Tal International Publishing Company 1976
Priess, H., Koller, B., Hess, B., Delius, H.: Electron microscopic mapping and sequence analysis of the terminator for the early message of E. coli phage T7. Mol. Gen. Genet. 178, 27–34 (1980)
Thomas, J.R., Tewari, K.K.: Conservation of 70S ribosomal RNA genes in the chloroplast DNAs of higher plants. Proc. Natl. Acad. Sci. U.S.A. 71, 3147–3151 (1974a)
Thomas, J.R., Tewari, K.K.: Ribosomal RNA genes in the chloroplast DNA of pea leaves. Biochim. Biophys. Acta 361, 73–83 (1974b)
Vedel, F., Quetier, F., Bayen, M.: Specific cleavage of chloroplast DNA from higher plants by EcoRI restriction nuclease. Nature 236, 440–442 (1976)
Whitfeld, P.R., Herrmann, R.G., Bottomley, W.: Mapping of the ribosomal RNA genes on chloroplast DNA. Nucleic Acids Res. 5, 1741–1751 (1978)
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Koller, B., Delius, H. Vicia faba chloroplast DNA has only one set of ribosomal RNA genes as shown by partial denaturation mapping and R-loop analysis. Molec. Gen. Genet. 178, 261–269 (1980). https://doi.org/10.1007/BF00270471
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DOI: https://doi.org/10.1007/BF00270471