Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1989 Oct;9(10):4398–4401. doi: 10.1128/mcb.9.10.4398

Microinjection of plasmid DNA encoding the A surface antigen of Paramecium tetraurelia restores the ability to regenerate a wild-type macronucleus.

S Koizumi 1, S Kobayashi 1
PMCID: PMC362521  PMID: 2586515

Abstract

Strain d48 of Paramecium tetraurelia contains the A i-antigen gene in the micronucleus, but the gene is lost when micronuclear products develop into the macronucleus. It has recently been shown that when injected into d48, macronucleoplasm from the wild type transforms d48 cells to wild type. It is shown here that wild-type cytoplasm can also bring about transformation, with a marked stage-specific sensitivity for both donor and recipient. It was also found that a plasmid containing the cloned A gene could transform d48 to wild type. Injection of nucleoplasm from animals in the vegetative stage of the cell cycle into the cytoplasm of recipients at various stages of autogamy caused high-frequency transformation of cells able to express the A serotype both before and after the next autogamy. Injection of nucleoplasm into vegetative macronuclei produced over 70% transformants able to express the A serotype after the next autogamy. The ability of nucleoplasm to transform was acquired at the second cell cycle after autogamy and was maintained throughout the vegetative stage. When cytoplasm was obtained from donors during autogamy and injected into the cytoplasm of recipients 1 to 2 h after the sensitive period, quite high frequencies of stable revertants were found when tested both before and after the next autogamy. Cells that were injected into the macronucleus with the cloned A plasmid expressed the A serotype after five fissions in over 20% of the lines and maintained this ability through successive fissions; all transformants except one stably expressed the A serotype even after the next autogamy.

Full text

PDF
4398

Selected References

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

  1. Berger J. D. Nuclear differentiation and nucleic acid synthesis in well-fed exconjugants of Paramecium aurelia. Chromosoma. 1973 Jun 28;42(3):247–268. doi: 10.1007/BF00284774. [DOI] [PubMed] [Google Scholar]
  2. Brygoo Y., Sonneborn T. M., Keller A. M., Dippell R. V., Schneller M. V. Genetic Analysis of Mating Type Differentiation in PARAMECIUM TETRAURELIA. II. Role of the Micronuclei in Mating-Type Determination. Genetics. 1980 Apr;94(4):951–959. doi: 10.1093/genetics/94.4.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Epstein L. M., Forney J. D. Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia. Mol Cell Biol. 1984 Aug;4(8):1583–1590. doi: 10.1128/mcb.4.8.1583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Godiska R., Aufderheide K. J., Gilley D., Hendrie P., Fitzwater T., Preer L. B., Polisky B., Preer J. R., Jr Transformation of Paramecium by microinjection of a cloned serotype gene. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7590–7594. doi: 10.1073/pnas.84.21.7590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grandchamp S., Beisson J. Positional control of nuclear differentiation in Paramecium. Dev Biol. 1981 Jan 30;81(2):336–341. doi: 10.1016/0012-1606(81)90297-9. [DOI] [PubMed] [Google Scholar]
  6. Harumoto T. Induced change in a non-mendelian determinant by transplantation of macronucleoplasm in Paramecium tetraurelia. Mol Cell Biol. 1986 Oct;6(10):3498–3501. doi: 10.1128/mcb.6.10.3498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Koizumi S., Kobayashi S. A Study of Odd Mating-Type Determination Factor by Transfer of Macronuclear Karyoplasm in PARAMECIUM TETRAURELIA. Genetics. 1984 Jul;107(3):367–373. doi: 10.1093/genetics/107.3.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Koizumi S., Kobayashi S. The maintenance of kappa during macronuclear reorganization in Paramecium tetraurelia. J Cell Sci. 1978 Apr;30:187–191. doi: 10.1242/jcs.30.1.187. [DOI] [PubMed] [Google Scholar]
  9. Koizumi S. Microinjection and transfer of cytoplasm in Paramecium. Experiments on the transfer of kappa particles into cells at different stages. Exp Cell Res. 1974 Sep;88(1):74–78. doi: 10.1016/0014-4827(74)90619-3. [DOI] [PubMed] [Google Scholar]
  10. Preer J. R., Preer L. B., Rudman B. M. mRNAs for the immobilization antigens of Paramecium. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6776–6778. doi: 10.1073/pnas.78.11.6776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sonneborn T. M. Genetics of cellular differentiation: stable nuclear differentiation in eucaryotic unicells. Annu Rev Genet. 1977;11:349–367. doi: 10.1146/annurev.ge.11.120177.002025. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES