Summary
The restriction endonucleases SalGI and PstI have been used to construct a physical map of wheat ctDNA. The molecule was found to contain approximately 135 kbq, and in common with many other higher plant ctDNAs about 15% of the sequences are repeated in an inverted orientation. It was established by electron microscopy that, in wheat, each segment of the inverted repeat contains 21.0 kbp, and that the single copy regions separating the two repeated segments contain 12.8 kbp and 80.2 kbp. Blot hybridisation showed that one set of ribosomal genes is located in each segment of the inverted repeat region and the sizes of these genes were accurately determined by measuring the dimensions of hybrids between the chloroplast rRNAs and the identified Sal and Eco fragments on electron micrographs: the genes for the 16S and 23S rRNAs contain 1530 bp and 2850 bp respectively and are separated by a spacer region of 2350 bp. The Bgl fragment of maize ctDNA known to contain the structural gene for the large-subunit (LS) of ribulose 1,5-bisphosphate carboxylase was used as a probe to locate the LS gene in wheat ctDNA. A small (2.8 kbp) Eco fragment was found to contain most of the wheat LS gene and is derived from the larger single-copy region, 23.5 kbp away from one segment of the inverted repeat and 54.8 kbp from the other.
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References
Bedbrook JR, Bogorad L (1976) Endonuclease recognition sites mapped on Zea mays chloroplast DNA. Proc Natl Acad Sci USA 73:4309–4313
Bedbrook JR, Kolodner R, Bogorad L (1977) Zea mays chloroplast ribosomal RNA genes are part of a 22000 base pair inverted repeat. Cell 11:739–749
Bedbrook JR, Coen DM, Beaton AR, Bogorad L, Rich A (1979) Location of the single gene for the large subunit of ribulose bisphosphate carboxylase on the maize chloroplast chromosome. J Biol Chem 254:3264–3271
Bedbrook JR, Kolodner R (1979) The structure of chloroplast DNA. Annu Rev Plant Physiol 30:593–620
Bogorad L, Jolly SO, Kidd G, Link G, McIntosh L (1980) Organisation and transcription of maize chloroplast genes. In: Leaver CJ (ed) Genome organisation and expression in plants. Plenum Press, New York, pp 291–304
Brosius J, Dull TJ, Noller HF (1980) Complete nucleotide sequence of a 23S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci USA 77:201–204
Chow LT, Roberts JM, Lewis B, Broker TR (1977) A map of cytoplasmic RNA transcripts from lytic adenovirus type 2 determined by electron microscopy of RNA DNA hybrids. Cell 11:819–836
Davis RW, Simon M, Davidson N (1971) Electron microscope heteroduplex methods for mapping regions of base sequence homology. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 21. Academic Press, New York, pp 413–428
Delius H, Koller B (1980) Sequence homologies between E. coli and chloroplast ribosomal DNA, as seen by heteroduplex analysis. J Mol Biol 142:247–261
Dyer TA, Bowman CM (1979) Nucleotide sequences of chloroplast 5S ribosomal ribonucleic acid in flowering plants. Biochem J 183:595–605
Gatenby AA, Castleton JA, Saul MW (1981) Expression in E. coli of the maize and wheat chloroplast genes for the large subunit set of ribosomal RNA genes as shown by partial denaturation mapping and R-loop analysis. Mol Gen Genet 178:261–269
Kolodner R, Tewari KK (1975) The molecular size and conformation of the chloroplast DNA from higher plants. Biochim Biophys Acta 402:372–390
Kolodner R, Tewari KK (1979) Inverted repeats in chloroplast DNA from higher plants. Proc Natl Acad Sci USA 76:41–45
Kusuda J, Shinozaki K, Takaiwa F, Sugiura M (1980) Characterization of the cloned ribosomal DNA on tobacco chloroplasts. Mol Gen Genet 178:1–7
Maizels N (1976) Dictyostelium 17S, 25S and 5S rDNAs lie within a 38000 base pair repeated unit. Cell 9:431–438
Portmann R, Koller Th (1976) 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
Priess H, Koller B, Hess B, Delius H (1980) Electron microscopic mapping and sequence analysis of the terminator for the early message of E. coli phage T7. Mol Gen Genet 178:27–34
Quetier F, Vedel F (1980) Physico-chemical restriction endonuclease analysis of mitochondrial DNA from higher plants. In: Leaver CJ (ed) Genome Organisation and Expression in Plants. Plenum Press, New York, pp 401–406
Rochaix JD, Malnoë P (1978) Anatomy of the chloroplast ribosomal DNA of Chlamydomonas reinhardii. Cell 15:661–670
Sanger F, Coulson AR, Barrel BG, Smith AGH, Roe BA (1980) Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178
Schwarz Z, Kössel H (1980) The primary structure of 16S rRNA from Zea mays chloroplasts is homologous to E. coli 16S rRNA.
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Bowman, C.M., Koller, B., Delius, H. et al. A physical map of wheat chloroplast DNA showing the location of the structural genes for the ribosomal RNAs and the large subunit of ribulose 1,5-bisphosphate carboxylase. Molec. Gen. Genet. 183, 93–101 (1981). https://doi.org/10.1007/BF00270145
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DOI: https://doi.org/10.1007/BF00270145