Abstract
We report the molecular cloning of the tomato golden mosaic virus (TGMV) genome in the E. coli plasmid pAT 153. The results of this work conclusively show that TGMV DNA consists of two components (designated A and B) of almost, but not exactly, the same size. Four different recombinant plasmids are described, two containing component A in opposite orientation and two containing component B in opposite orientation. Southern blot analysis revealed little sequence homology between A and B and showed both components to be equally represented in viral and intracellular DNA forms. Detailed restriction maps of the cloned DNAs are presented, and a comparison of these with digests of intracellular viral dsDNA indicates that the former are full-length faithful copies of TGMV DNA. This is the first report of the cloning of a geminivirus genome.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Buck K. W., Ratti G. Molecular weight of double-stranded RNA: a re-examination of Aspergillus foetidus virus S RNA components. J Gen Virol. 1977 Oct;37(1):215–219. doi: 10.1099/0022-1317-37-1-215. [DOI] [PubMed] [Google Scholar]
- Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
- Efstratiadis A., Vournakis J. N., Donis-Keller H., Chaconas G., Dougall D. K., Kafatos F. C. End labeling of enzymatically decapped mRNA. Nucleic Acids Res. 1977 Dec;4(12):4165–4174. doi: 10.1093/nar/4.12.4165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Schuerch A. R., Mitchell W. R., Joklik W. K. Isolation of intact individual species of single- and double-stranded RNA after fractionation by polyacrylamide gel electrophoresis. Anal Biochem. 1975 May 12;65(1-2):331–345. doi: 10.1016/0003-2697(75)90517-5. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Sänger H. L. Functions of the two particles of tobacco rattle virus. J Virol. 1969 Mar;3(3):304–312. doi: 10.1128/jvi.3.3.304-312.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
- Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]