Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1991 May;96(1):280–284. doi: 10.1104/pp.96.1.280

Cloning and Inactivation of a Gene Essential to Inorganic Carbon Transport of Synechocystis PCC6803 1

Teruo Ogawa 1
PMCID: PMC1080746  PMID: 16668165

Abstract

A clone (HP-1) which transforms the high CO2-requiring mutant (RKb) of Synechocystis PCC6803 defective in inorganic carbon transport to the wild-type (WT) phenotype was isolated from a WT genomic library. The clone contained a 5.4 kilobase-pair DNA insert. Complementation tests with subclones derived from HP-1 allowed the mutation in RKb to be located within 141 base-pair nucleotides. Sequencing of nucleotides in this region revealed an open reading frame encoding a hydrophobic protein consists of 80 amino acids. A defined mutant (M9) constructed by inactivating this putative inorganic carbon transport gene, designated ictA, was unable to transport CO2 and HCO3− into the intracellular inorganic carbon pool. Cloning and sequence analysis of the respective RKb gene revealed a base substitution which generates a stop codon in the middle of ictA.

Full text

PDF
281

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Badger M. R., Kaplan A., Berry J. A. Internal Inorganic Carbon Pool of Chlamydomonas reinhardtii: EVIDENCE FOR A CARBON DIOXIDE-CONCENTRATING MECHANISM. Plant Physiol. 1980 Sep;66(3):407–413. doi: 10.1104/pp.66.3.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dzelzkalns V. A., Bogorad L. Molecular analysis of a mutant defective in photosynthetic oxygen evolution and isolation of a complementing clone by a novel screening procedure. EMBO J. 1988 Feb;7(2):333–338. doi: 10.1002/j.1460-2075.1988.tb02817.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Espie G. S., Miller A. G., Birch D. G., Canvin D. T. Simultaneous Transport of CO(2) and HCO(3) by the Cyanobacterium Synechococcus UTEX 625. Plant Physiol. 1988 Jul;87(3):551–554. doi: 10.1104/pp.87.3.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Espie G. S., Miller A. G., Canvin D. T. Characterization of the na-requirement in cyanobacterial photosynthesis. Plant Physiol. 1988 Nov;88(3):757–763. doi: 10.1104/pp.88.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kaplan A., Zenvirth D., Marcus Y., Omata T., Ogawa T. Energization and activation of inorganic carbon uptake by light in cyanobacteria. Plant Physiol. 1987 Jun;84(2):210–213. doi: 10.1104/pp.84.2.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  7. Miller A. G., Colman B. Active transport and accumulation of bicarbonate by a unicellular cyanobacterium. J Bacteriol. 1980 Sep;143(3):1253–1259. doi: 10.1128/jb.143.3.1253-1259.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ogawa T., Kaplan A. The Stoichiometry between CO(2) and H Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria. Plant Physiol. 1987 Apr;83(4):888–891. doi: 10.1104/pp.83.4.888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ogawa T. Mutants of Synechocystis PCC6803 Defective in Inorganic Carbon Transport. Plant Physiol. 1990 Oct;94(2):760–765. doi: 10.1104/pp.94.2.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Oka A., Sugisaki H., Takanami M. Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol. 1981 Apr 5;147(2):217–226. doi: 10.1016/0022-2836(81)90438-1. [DOI] [PubMed] [Google Scholar]
  11. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Price G. D., Badger M. R. Ethoxyzolamide Inhibition of CO(2) Uptake in the Cyanobacterium Synechococcus PCC7942 without Apparent Inhibition of Internal Carbonic Anhydrase Activity. Plant Physiol. 1989 Jan;89(1):37–43. doi: 10.1104/pp.89.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  14. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  15. Stanier R. Y., Kunisawa R., Mandel M., Cohen-Bazire G. Purification and properties of unicellular blue-green algae (order Chroococcales). Bacteriol Rev. 1971 Jun;35(2):171–205. doi: 10.1128/br.35.2.171-205.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Volokita M., Zenvirth D., Kaplan A., Reinhold L. Nature of the Inorganic Carbon Species Actively Taken Up by the Cyanobacterium Anabaena variabilis. Plant Physiol. 1984 Nov;76(3):599–602. doi: 10.1104/pp.76.3.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Williams J. G., Szalay A. A. Stable integration of foreign DNA into the chromosome of the cyanobacterium Synechococcus R2. Gene. 1983 Sep;24(1):37–51. doi: 10.1016/0378-1119(83)90129-4. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES