Construction of Specific Mutations in Photosystem II Photosynthetic Reaction Center by Genetic Engineering Methods in Synechocystis 6803

Methods in Enzymology

Volumn 167, Issue C, 1988, Pages 766-778

  Williams, John G K ^a  

^a NONE

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EID: 77957024978     PISSN: 00766879     EISSN: 15577988     Source Type: Book Series    
DOI: 10.1016/0076-6879(88)67088-1     Document Type: Article

References (24)

1. Griogorieva, G., Shestakov, S., FEMS Microbiol. Lett., 13, 1982, 367.
2. Rippka, R., Arch Mikrobiol., 87, 1972, 93 In photoheterotrophic metabolism, both glucose and light are needed for growth when PSII is inactivated. The action spectrum for photoheterotrophic growth of Synechocystis 6803 is the absorption spectrum for chlorophyll plus a minor contribution from phycobilin [W. F. J. Vermaas and T. Ogawa, unpublished observations (1986)]. This indicates that photosystem I (PSI) is needed for photoheterotrophic growth, probably to generate ATP through cyclic electron transport. In this regard, we have found that Synechocystis 6803 grows very slowly on glycose in the dark, with a doubling time of 84 hr. Therefore, PSI is not absolutely required for growth but it is important for achieving practical growth rates. We believe that PSI is important, even in the presence of glucose, and that it is therefore unavailable to modification by the methods described here for PSII.
3. Pakrasi, H.B., Reithma, H.C., Sherman, L.A., Proc. Natl. Acad.Sci. U.S.A., 82, 1985, 6903.
4. Arntzen, C.J., Pakrasi, H.B., Encycl. Plant Physiol., New Ser., 19, 1986, 457.
5. Tang, X.-S., Satoh, K., FEBS Lett., 201, 1986, 221.
6. Vermaas, W.F.J., Williams, J.G.K., Rutherford, A.W., Mathis, P., Arntzen, C.J., Proc. Natl. Acad. Sci. U.S.A., 83, 1986, 4974.
7. This chapter, and also footnote h in Table I.
8. R.J. Rothstein, this series, Vol. 101, p. 202.
9. Wiliams, J.G.K., Szalay, A.A., Gene, 24, 1983, 37.
10. Kolowsky, K.S., Williams, J.G.K., Szalay, A.A., Gene, 27, 1984, 289.
11. D.A. Bryant, personal communication (1986).
12. Folters were Rec-85 membranes, 85 mm diameter, presterilized, Stock Number 145318, from Nuclepore Corp., 7035 Commerce Circle, Pleasanton, CA 94566-3294. Nitrocellulose membranes from Millipore or Schleicher & Schuell have also been used successfully.
13. B. Haskell and L.A. Sherman, personal communication (1986).
14. Hauska, G., Encycl. Plannt Physiol., New Ser., 5, 1977, 255.
15. Vermaas, W.F.J., Williams, J.G.K., Arntzen, C.J., Z. Naturforsch. C, 42, 1987, 128.
16. Debus, R.J., Barry, B.A., Babcock, G.T., McIntosh, L., Proc. Natl. Acad. Sci. U.S.A., 85, 1988, 427.
17. Danner, D.B., Smith, H.O., Narang, S.A., Proc. Natl. Acad. Sci. U.S.A., 79, 1982, 2393.
18. If donor DNA were clipped into smaller segments during transport as occurs in naturally transformable bacteria [M.L. Pifer and H.O. Smith, Proc. Natl. Acad. Sci. U.S.A. 82, 3731 (1985)], then overlapping fragments could be pieced together to regenerate an intact donor molecule capable of integration; overlapping fragments could only be produced if two or more donor molecules were transported into a single cell.
19. Fructose is toxic to the original strain of Synechococcus 6803 obtained from Dr. C. P. Wolk [E. Flores and G. Schmetterer, J. Bacteriol. 166, 693 (1986)]. The new strain, which was selected for glucose tolerance, was found also to be fructose resistant. Fructose is not utilized for growth by either the original strain or the new strain. An impairment in sugar transport has been ruled out as an explanation for sugar tolerance in the new strain, since it transports glucose normally [G. Schmetterer and J. G. K. Williams, unpublished observations (1986)].
20. Rippka, R., Deruelles, J., Waterbury, J.B., Herdman, M., Stanier, R.Y., J. Gen. Microbiol., 111, 1979, 1.
21. Utkilen, H.C., J. Gen. Microbiol, 95, 1976, 177.
22. Transformation-defective variants of another cyanobacterium, Synechococcus R2, have been isolated fortuitously by picking single colonies in the course of culture maintenance [J.G.K. Williams and K.S. Kolowsky, unpublished observations (1983)].
23. Synechocystis 6803 is notoriously resistant to digestion with lysozyme. Pretreatment of cells with 4% sodium dodecyl sulfate (SDS) at 75° for 15 min, or with saturated NaI as described above, allows lysozyme to digest the cells wall. These procedures are throught to remove a protective coating from the cell surface. NaI is preferred over SDS because it is easier to wash way from the cells.
24. Maniatis, T., Fritsch, E.F., Sambrook, J., Molecular Cloning: A Laboratory Manual, 1982, Cold Spring Harbor Lab, Cold Spring Harbor, New York, 353.