Mages, Wolfgang and Heinrich, Oliver and Treuner, Gerda and Vlcek, Daniel and Daubnerova, Ivana and Slaninova, Miroslava (2007) Complementation of the Chlamydomonas reinhardtii arg7-8 (arg2) point mutation by recombination with a truncated nonfunctional ARG7 gene. PROTIST, 158 (4). pp. 435-446. ISSN 1434-4610,
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Chlamydomonas reinhardtii arg7-8 (arg2) mutant strains carrying a hitherto undescribed mutation in their argininosuccinate lyase gene ( ARG7) that leads to arginine auxotrophy have been used together with the corresponding wild-type gene as a very reliable transformation system since 1989. In this study, we finally identify the molecular nature of the arg7-8 mutation as a (6073)G to A transition in exon 9 of ARG7 leading to a (288)Gly to Ser exchange near the active site of the protein. The same mutation was found in the ARG7 genes of three commonly used C. reinhardtii laboratory strains, namely cw 15-302 arg2, CC-48, and CC-1618. We did not observe exact spontaneous reversion of the arg7-8 allele in our study, but did identify two different and rare intragenic suppressor mutations, (27)Leu to Phe and (285)Tyr to Phe. In our hands, only transformation of the arg7-8 strain with a truncated nonfunctional wild-type ARG7 gene lacking 124 codons at its 50 end led to exact reversion of the mutant base (6073)A to the wildtype (6073)G, presumably by recombination. This system offers a positive selection scheme for homologous recombination (HR) and may, therefore, be useful to the methodical improvement of recombination in Chlamydomonas. (c) 2007 Elsevier GmbH. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ARGININOSUCCINATE LYASE GENE; SINGLE-STRANDED-DNA; HOMOLOGOUS RECOMBINATION; NUCLEAR TRANSFORMATION; INTRAGENIC COMPLEMENTATION; INTRAFLAGELLAR TRANSPORT; INSERTIONAL MUTAGENESIS; RESISTANCE; MUTANTS; CLONING; intragenic suppressor; nuclear transformation; marker; targeting |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Cell Cycle Control |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 01 Dec 2020 13:06 |
| Last Modified: | 01 Dec 2020 13:06 |
| URI: | https://pred.uni-regensburg.de/id/eprint/32064 |
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