1887

Abstract

Summary

A simple and rapid labelling technique is described for preparing tobacco mosaic virus labelled with I. The technique is versatile and has been used to obtain labelled virus of both high and low specific activity. High specific activity virus has been used in a pilot radioautographic experiment at the electron microscope level, and micrographs were obtained showing virus particles in close association with silver grains. At specific activities of 2 to 3 mc/mg. a rapid loss of infectivity was observed after labelling; this effect was not observed when the specific activity was 1 c/mg. The sensitivity of detection for high specific activity preparations by scintillation counting and by electron microscope radioautography is discussed and the potentiality of I-labelled virus of both high and low specific activity is indicated.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-2-1-89
1968-01-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/2/1/JV0020010089.html?itemId=/content/journal/jgv/10.1099/0022-1317-2-1-89&mimeType=html&fmt=ahah

References

  1. Ada G. L., Humphrey J. H., Askonas B. A., Mcdevtit H. O., Nossal G. J. V. 1966; Correlation of grain counts with radioactivity (125I and tritium) in autoradiography. Expl Cell Res 41:557
    [Google Scholar]
  2. Anson M. L., Stanley W. M. 1941; Some effects of iodine and other reagents on the structure and activity of tobacco mosaic virus. J. gen. Physiol 24:679
    [Google Scholar]
  3. Appelgren L., Soremark R., Ullberg S. 1963; Improved resolution in autoradiography with radioiodine using the extranuclear electron radiation from 126I. Biochim. biophys. Acta 66:144
    [Google Scholar]
  4. Bachmann L., Salpeter M. M. 1965; Autoradiography with the electron microscope, a quantitative evaluation. Lab. Invest 14:1041
    [Google Scholar]
  5. Brants D. H. 1963; Transport of 14C-labeIled tobacco mosaic virus material in tobacco leaves. Virology ao:388
    [Google Scholar]
  6. Brants D. H. 1965; Relation between ectodesmata and infection of leaves by 14C-labelled tobacco mosaic virus. Virology 26:554
    [Google Scholar]
  7. Cairns J. 1962; The application of autoradiography to the study of DNA viruses. Cold Spring Harb. Symp. quant. Biol 27:311
    [Google Scholar]
  8. Caro L. G. 1962; High resolution autoradiography. II. The problem of resolution. J. Cell Biol 15:189
    [Google Scholar]
  9. Cocking E. C. 1966; An electron microscopic study of the initial stages of infection of isolated tomato fruit protoplasts by tobacco mosaic virus. Planta 68:206
    [Google Scholar]
  10. Dales S. 1963; The uptake and development of vaccinia virus in strain L cells followed with labelled viral DNA. J. Cell Biol 18:51
    [Google Scholar]
  11. De Wachter R., Fiers W. 1967; External standardization in liquid scintillation counting of homogeneous samples labelled with one, two or three isotopes. Analyt. Biochem 18:351
    [Google Scholar]
  12. Fraenkel-Conrat H. 1955; The reaction of tobacco mosaic virus with iodine. J. biol. Chem 217:373
    [Google Scholar]
  13. Gregory D. W., Cocking E. C. 1965; The large scale isolation of protoplasts from immature tomato fruit. J. Cell Biol 24:143
    [Google Scholar]
  14. Hall T. C., Cocking E. C. 1965; High-efficiency liquid-scintillation counting of 14C-Iabelled material in aqueous solution and determination of specific activity of labelled proteins. Biochem. J 96:626
    [Google Scholar]
  15. Herridge E. A., Schlegel D. E. 1962; Autoradiographic studies of tobacco mosaic virus inoculations on host and non-host species. Virology 18:517
    [Google Scholar]
  16. Kontaxis D. G., Schlegel D. E. 1962; Basal septa of broken trichomes in Nicotiana as possible infection sites for tobacco mosaic virus. Virology 16:244
    [Google Scholar]
  17. Maunsbach A. B. 1966; Absorption of 125I labelled homologous albumin by rat kidney proximal tubule ceils. A study of microperfused single proximal tubules by electron microscopic autoradiography and histochemistry. J. Ultrastruct. Res 15:197
    [Google Scholar]
  18. Mohr W., Cocking E. C. 1967; A method for preparing highly vacuolated, senescent and damaged plant tissue for ultrastructural studies. J. Ultrastruct. Res In the Press
    [Google Scholar]
  19. Reynolds E. S. 1963; The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol 17:208
    [Google Scholar]
  20. Rhodes B. A. 1965; Liquid scintillation counting of radioiodine. Analyt. Chem 37:995
    [Google Scholar]
  21. Ryser H. J. P. 1967; Studies on protein uptake by isolated tumor cells. III. J. Cell Biol 32:737
    [Google Scholar]
  22. Salpeter M. M., Bachmann L. 1964; Autoradiography with the electron microscope. A procedure for improving resolution, sensitivity and contrast. J. Cell Biol 22:469
    [Google Scholar]
  23. Salpeter M. M., Bachmann L. 1965; Assessment of technical steps in electron microscope autoradiography. Symp. bit. Soc. Cell Biol 4:23 in discussion
    [Google Scholar]
  24. Stevens A. R. 1966; High resolution autoradiography. In Methods in Cell Physiology 2255 Prescott D. M. Ed. by New York: Academic Press;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-2-1-89
Loading
/content/journal/jgv/10.1099/0022-1317-2-1-89
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error