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Construction and mapping of safflower chloroplast DNA recombinants and location of selected gene markers

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Summary

DNA was isolated and purified from chloroplasts of safflower (Carthamus tinctorius L.), digested with HindIII restriction endonuclease, and ligated into the HindIII site of the plasmid pUC9. Recombinant DNAs were isolated from ampicillin resistant white colonies which grew in the presence of the appropriate indicator, digested with HindIII, and then identified by comparison of agarose gel electrophoretic mobilities. HindIII digests of chloroplast DNA were used as a standard. Such recombinants were radiolabeled and hybridized with Southern blots of PstI, SalI, KpnI, and HindIII single and double digests of safflower chloroplast DNA. A physical map was subsequently generated showing the location of each recombinant on the circular plastid genome. Recombinants containing heterologous chloroplast gene markers from spinach or Euglena were also radiolabeled and mapped. The relative mapping positions of these genes are in good agreement with those which have previously been published for spinach and several other higher plants.

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References

  1. Bohnert HJ, Crouse EJ, Schmidt JM (1982) Nucleic acids and proteins in plants. In: Boulter D, Parthier D (eds) Encyclopedia of plant physiology, vol 14 B. Springer, Berlin Heidelberg Stuttgart, pp 475–530

  2. Edelman M (1981) Nucleic acids of chloroplasts and mitochondria. In: Stumf PK, Conn EE (eds) The biochemistry of plants, vol 6. Academic Press, New York, pp 249–301

  3. Fluhr R, Fromm H, Edelman M (1983) Clone bank of Nicotiana tobacum chloroplast DNA: mapping of the alpha, beta and epsilon subunits of the ATPase coupling factor, the large subunit of ribulosebisphosphate carboxylase, and the 32-kDal membrane protein. Gene 25:271–280

  4. Hollingsworth MJ, Johanningmeier U, Karabin GD, Stiegler GL, Hallick RB (1984) Detection of multiple, unspliced precursor mRNA transcripts for the Mr 32,000 thylakoid membrane protein from Euglena gracilis chloroplasts. Nucleic Acid Res 12:2001–2017

  5. Ma C, Tyrawska D, Smith MA (1984) Isolation and characterization of safflower (Carthamus tinctorius L.) chloroplast DNA. Mol Cell Biochem 65:181–188

  6. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a labroatory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

  7. Overbeeke N, Haring MA, John H, Nijkamp J, Kool AJ (1984) Cloning of Petunia hybrida chloroplast DNA sequences capable of autonomous replication in yeast. Plant Mol Biol 3:235–241

  8. Palmer JD, Thompson WF (1981) Clone banks of the mung bean, pea and spinach chloroplast genomes. Gene 15:21–26

  9. Palmer JD, Thompson WF (1982) Chloroplast DNA rearrangements are more frequent when a large inverted repeat sequence is lost. Cell 29:537–550

  10. Smith GE, Summers MD (1980) The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethol-paper. Anal Biochem 109:123–129

  11. Uchimiya H, Ohtani T, Ohgawara T, Harada H, Sugita M, Suguira M (1983) Molecular cloning of tobacco chromosomal and chloroplast DNA segments capable of replication in yeast. Mol Gen Genet 192:1–4

  12. Vedel F, Mathieu C (1983) Physical mapping of chloroplast DNA from normal and cytoplasmic male sterile (radish cytoplasm) lines of Brassica napus. Curr Genet 7:13–20

  13. Weil JH, Mubumbila M, Kuntz M, Keller M, Steinmetz A, Crouse EJ, Burkard G, Gillemant P, Selden R, McIntosh L, Bogorad L, Loffelhardt W, Mucke H, Bohnert HJ (1983) Gene mapping studies and sequence determination of chloroplast transfer RNAs from various photosynthetic organisms. Mol Biol (Moskow) 17:1147–1153

  14. Whitfeld PR, Bottomley W (1983) Organization and structure of chloroplast genes. Annu Rev Plant Physiol 34:279–310

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Author information

Correspondence to M. A. Smith.

Additional information

Communicated by D. von Wettstein

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Ma, C., Smith, M.A. Construction and mapping of safflower chloroplast DNA recombinants and location of selected gene markers. Theoret. Appl. Genetics 70, 620–627 (1985). https://doi.org/10.1007/BF00252287

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Key words

  • Safflower
  • Chloroplast
  • DNA
  • Clone-bank
  • Mapping