Abstract
Carrier-free iodination of 5S RNA with 125I yields a probe suitable for use in cytological RNA-DNA hybridization studies. The genes coding for 5S ribosomal RNA in Drosophila could be localized by autoradiography after a 2-day exposure, whereas a 2-month exposure is needed when the best available [3H]RNA is used. The procedure introduces one covalently bound 125I atom per 100 nucleotides, resulting in a specific activity of over 108 dpm/μg of RNA. The method may be readily applied to other problems involving molecular hybridization techniques.
Keywords: molecular hybridization, chromosomes, iodine-125
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ada G. L., Humphrey J. H., Askonas B. A., McDevitt H. O., Nossal G. J. Correlation of grain counts with radioactivity (125I and tritium) in autoradiography. Exp Cell Res. 1966 Mar;41(3):557–572. doi: 10.1016/s0014-4827(66)80106-4. [DOI] [PubMed] [Google Scholar]
- Commerford S. L. Iodination of nucleic acids in vitro. Biochemistry. 1971 May 25;10(11):1993–2000. doi: 10.1021/bi00787a005. [DOI] [PubMed] [Google Scholar]
- Ertl H. H., Feinendegen L. E., Heiniger H. J. Iodine-125, a tracer in cell biology: physical properties and biological aspects. Phys Med Biol. 1970 Jul;15(3):447–456. doi: 10.1088/0031-9155/15/3/005. [DOI] [PubMed] [Google Scholar]
- Getz M. J., Altenburg L. C., Saunders G. F. The use of RNA labeled in vitro with iodine-125 in molecular hybridization experiments. Biochim Biophys Acta. 1972 Dec 22;287(3):485–494. doi: 10.1016/0005-2787(72)90293-6. [DOI] [PubMed] [Google Scholar]
- Henderson A. S., Warburton D., Atwood K. C. Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3394–3398. doi: 10.1073/pnas.69.11.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henderson A. S., Warburton D., Atwood K. C. Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3394–3398. doi: 10.1073/pnas.69.11.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones K. W. Chromosomal and nuclear location of mouse satellite DNA in individual cells. Nature. 1970 Mar 7;225(5236):912–915. doi: 10.1038/225912a0. [DOI] [PubMed] [Google Scholar]
- Lambert B., Wieslander L., Daneholt B., Egyházi E., Ringborg U. In situ demonstration of DNA hybridizing with chromosomal and nuclear sap RNA in Chironomus tentans. J Cell Biol. 1972 May;53(2):407–418. doi: 10.1083/jcb.53.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardu M. L., Gerbi S. A., Eckhardt R. A., Gall J. G. Cytological localization of DNA complementary to ribosomal RNA in polytene chromosomes of Diptera. Chromosoma. 1970;29(3):268–290. doi: 10.1007/BF00325943. [DOI] [PubMed] [Google Scholar]
- Pardue M. L., Gall J. G. Chromosomal localization of mouse satellite DNA. Science. 1970 Jun 12;168(3937):1356–1358. doi: 10.1126/science.168.3937.1356. [DOI] [PubMed] [Google Scholar]
- Pardue M. L., Gall J. G. Molecular hybridization of radioactive DNA to the DNA of cytological preparations. Proc Natl Acad Sci U S A. 1969 Oct;64(2):600–604. doi: 10.1073/pnas.64.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steffensen D. M., Wimber D. E. Localization of tRNA genes in the salivary chromosomes of Drosophila by RNA:DNA hybridization. Genetics. 1971 Oct;69(2):163–178. doi: 10.1093/genetics/69.2.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wimber D. E., Steffensen D. M. Localization of 5S RNA genes on Drosophila chromosomes by RNA-DNA hybridization. Science. 1970 Nov 6;170(3958):639–641. doi: 10.1126/science.170.3958.639. [DOI] [PubMed] [Google Scholar]