Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Dec;116(3):1170–1176. doi: 10.1128/jb.116.3.1170-1176.1973

Growth and Cloning of Tetrahymena pyriformis on Solid Medium

Enore Gardonio 1, Michael Crerar 1, Ronald E Pearlman 1
PMCID: PMC246471  PMID: 4127626

Abstract

A method of obtaining clones of Tetrahymena pyriformis on solid medium has been developed. The medium consists of a basal layer of 1.5% agar topped with 2 ml of 0.3% agar in sterile, plastic petri plates (100 by 15 mm). Both agar layers contain either 2% proteose peptone and 0.1% liver extract (complex medium) or defined medium supplemented with proteose peptone. After drying, 0.5 ml of liquid culture is spread evenly over the top agar, and the plates are then sprinkled lightly and evenly with autoclaved dry Sephadex G-25 (fine). Cell colonies can be observed after 5 days of incubation either by viewing with a microscope or without the aid of a microscope after staining. Plating efficiency is high on either complex or defined medium with a number of strains of Tetrahymena, both micronucleate and amicronucleate. Colonies can be picked and transferred to liquid culture for further growth. The existence of clones was demonstrated by plating a mixture of two different drug-resistant mutants. The method should prove useful in selective procedures for the isolation of mutants and for determining survival after treatments such as ultraviolet irradiation.

Full text

PDF
1170

Images in this article

1171Image
on p.1171
1172Image
on p.1172

Selected References

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

  1. Brunk C. F. The time sequence of events during excision repair in Tetrahymena pyriformis. Photochem Photobiol. 1972 Apr;15(4):349–360. doi: 10.1111/j.1751-1097.1972.tb06245.x. [DOI] [PubMed] [Google Scholar]
  2. CHRISTENSEN E., GIESE A. C. Increased photoreversal of ultraviolet injury by flashing light. J Gen Physiol. 1956 Mar 20;39(4):513–526. doi: 10.1085/jgp.39.4.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cameron I. L., Jeter J. R., Jr Synchronization of the cell cycle of Tetrahymena by stravation and refeeding. J Protozool. 1970 Aug;17(3):429–431. doi: 10.1111/j.1550-7408.1970.tb04708.x. [DOI] [PubMed] [Google Scholar]
  4. Crerar M., Pearlman R. E. DNA polymerase activity from Tetrahymena pyriformis. FEBS Lett. 1971 Nov 1;18(2):231–237. doi: 10.1016/0014-5793(71)80452-0. [DOI] [PubMed] [Google Scholar]
  5. HAMBURGER K., ZEUTHEN E. Synchronous divisions in Tetrahymena pyriformis as studied in an inorganic medium; the effect of 2,4-dinitrophenol. Exp Cell Res. 1957 Dec;13(3):443–453. doi: 10.1016/0014-4827(57)90074-5. [DOI] [PubMed] [Google Scholar]
  6. Hjelm K. K. A synchronizing effect of carbon dioxide on cell division in Tetrahymena. Exp Cell Res. 1971 Jul;67(1):204–207. doi: 10.1016/0014-4827(71)90635-5. [DOI] [PubMed] [Google Scholar]
  7. Kurtz S., Pearlman R. E. DNA-dependent RNA polymerase from Tetrahymena pyriformis. Can J Biochem. 1972 Feb;50(2):154–157. doi: 10.1139/o72-020. [DOI] [PubMed] [Google Scholar]
  8. PADILLA G. M., CAMERON I. L. SYNCHRONIZATION OF CELL DIVISION IN TETRAHYMENA PYRIFORMIS BY A REPETITIVE TEMPERATURE CYCLE. J Cell Physiol. 1964 Dec;64:303–307. doi: 10.1002/jcp.1030640304. [DOI] [PubMed] [Google Scholar]
  9. Roberts C. T., Jr, Orias E. Cytoplasmic inheritance of chloramphenicol resistance in tetrahymena. Genetics. 1973 Feb;73(2):259–272. doi: 10.1093/genetics/73.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. SCHERBAUM O., ZEUTHEN E. Induction of synchronous cell division in mass cultures of Tetrahymena piriformis. Exp Cell Res. 1954 Feb;6(1):221–227. doi: 10.1016/0014-4827(54)90164-0. [DOI] [PubMed] [Google Scholar]
  11. Sedgley N. N., Stone G. E. DNA synthesis in vinblastine synchronized Tetrahymena. Exp Cell Res. 1969 Jul;56(1):174–177. doi: 10.1016/0014-4827(69)90416-9. [DOI] [PubMed] [Google Scholar]
  12. Villadsen I. S., Zeuthen E. Synchronization of DNA synthesis in Tetrahymena populations by temporary limitation of access to thymine compounds. Exp Cell Res. 1970 Aug;61(2):302–310. doi: 10.1016/0014-4827(70)90452-0. [DOI] [PubMed] [Google Scholar]
  13. Wolfe J. Differential density labeling and gradient centrifugation of Tetrahymena: a new procedure for selection synchrony. Exp Cell Res. 1973 Mar 15;77(1):232–238. doi: 10.1016/0014-4827(73)90572-7. [DOI] [PubMed] [Google Scholar]
  14. Wunderlich F., Peyk D. Antimitotic agents and macronuclear division of ciliates. II. Endogenous recovery from colchicine and Colcemid--a new method of synchronization in Tetrahymena pyriformis GL. Exp Cell Res. 1969 Sep;57(1):142–144. doi: 10.1016/0014-4827(69)90379-6. [DOI] [PubMed] [Google Scholar]
  15. Zeuthen E. Synchrony in Tetrahymena by heat shocks spaced a normal cell generation apart. Exp Cell Res. 1971 Sep;68(1):49–60. doi: 10.1016/0014-4827(71)90585-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES