Babinger, Patrick and Kobl, Iris and Mages, Wolfgang and Schmitt, Ruediger (2001) A link between DNA methylation and epigenetic silencing in transgenic Volvox carteri. NUCLEIC ACIDS RESEARCH, 29 (6). pp. 1261-1271. ISSN 0305-1048
Full text not available from this repository. (Request a copy)Abstract
Epigenetic silencing of foreign genes introduced into plants poses an unsolved problem for transgenic technology. Here we have used the simple multicellular green alga Volvox carteri as a model to analyse the relation of DNA methylation to transgenic silencing. Volvox DNA contains on average 1.1% Ei-methylcytosine and 0.3% N6-methyladenine, as revealed by electrospray mass spectrometry and phosphoimaging of chromatographically separated P-32-labelled nucleotides. In two nuclear transformants of V.carteri, produced in 1993 by biolistic bombardment with a foreign arylsulphatase gene (C-ars), the transgene is still expressed in one (Hill 181), but not in the other (Hill 183), after an estimated 500-1000 generations. Each transformant clone contains multiple intact copies of C-ars, most of them integrated into the genome as tandem repeats. When the bisulphite genomic sequencing protocol was applied to examine two select regions of transgenic C-ars, we found that the inactivated copies (Hill 183) exhibited a high-level methylation (40%) of CpG dinucleotides, whereas the active copies (Hill 181) displayed low-level (7%) CpG methylation. These are average values from 40 PCR clones sequenced from each DNA strand in the two portions of C-ars. The observed correlation of CpG methylation and transgene inactivation in a green alga will be discussed in the light of transcriptional silencing.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DOMINANT SELECTABLE MARKER; CHLAMYDOMONAS-REINHARDTII; GENE-EXPRESSION; CHROMATIN STRUCTURE; MASS-SPECTROMETRY; NITRATE REDUCTASE; REPORTER-GENE; LIFE-CYCLE; PROTEIN; ARYLSULFATASE; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 08 Feb 2022 16:09 |
| Last Modified: | 08 Feb 2022 16:09 |
| URI: | https://pred.uni-regensburg.de/id/eprint/41661 |
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