Berthold, Peter and Schmitt, Rüdiger and Mages, Wolfgang (2002) An engineered Streptomyces hygroscopicus aph 7" gene mediates dominant resistance against hygromycin B in Chlamydomonas reinhardtii. PROTIST, 153 (4). pp. 401-412. ISSN 1434-4610
Full text not available from this repository.Abstract
We have developed a positively selectable marker for the green alga Chlamydomonas reinhardtii using the Streptomyces hygroscopicus aminoglycoside phosphotransferase gene (aph7"). Its expression is controlled by C. reinhardtii regulatory elements, namely, the beta2-tubulin gene promoter in combination with the first intron and the 3' untranslated region of the small subunit of ribulose bisphosphate carboxylase, rbcS2. C. reinhardtii cell-wall deficient and wild-type strains were transformed at rates up to 5 x 10(-5) with two constructs, pHyg3 and pHyg4 (intron-less). Transformants selected on plates with 10 mug/ml hygromycin B exhibited diverse levels of resistance of up to 200 mug/ml that were stably maintained for at least seven months; they contained two to five copies of the construct integrated in their genomes. Transcription of the chimeric aph7" gene, correct splicing of the rbcS2 intron, and polyadenylation of the transcripts have been verified by sequencing of RT-PCR products. Average co-transformation rates using pHyg3 and a second selectable plasmid were about 11%. This advocates the hygromycin-resistance plasmid, pHyg3, as a new versatile tool for the transformation of a broad range of C. reinhardtii strains without the sustained need for using auxotrophic mutants as recipients.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | NUCLEAR TRANSFORMATION; SELECTABLE MARKER; FOREIGN GENE; NITRATE REDUCTASE; EXPRESSION; VOLVOX; DNA; SEQUENCE; PROMOTER; PROTEIN; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Aug 2021 11:34 |
| Last Modified: | 26 Aug 2021 11:34 |
| URI: | https://pred.uni-regensburg.de/id/eprint/39643 |
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