Gruber, Heribert and Kirzinger, Stefan H. and Schmitt, Rüdiger (1996) Expression of the Volvox gene encoding nitrate reductase: Mutation-dependent activation of cryptic splice sites and intron-enhanced gene expression from a cDNA. PLANT MOLECULAR BIOLOGY, 31 (1). pp. 1-12. ISSN 0167-4412, 1573-5028
Full text not available from this repository.Abstract
Use of the nitrate reductase encoding gene (nitA) as selection marker has facilitated the successful nuclear transformation of Volvox carteri. The Volvox nitA gene contains 10 introns. A stable nitA mutation in the Volvox recipient strain 153-81 resides in a G-to-A transition of the first nucleotide in the 5' splice site of nitA intron 2. This mutation resulted in at least three non-functional splice variants, namely: (1) intron 2 was not spliced at all; (2) a cryptic 5' splice site 60 nt upstream or (3) a cryptic 5' splice site 16 nt downstream of the mutation were activated and used for splicing. When we used nitA cDNA (pVcNR13) for transformation of V. carteri 153-81, a low efficiency of about 5 x 10(-5) transformants per reproductive cell was observed. Re-integration of either intron 1 (pVcNR15) or introns 9 and 10 (pVcNR16) in the transforming cDNA increased transformation rates to 5 x 10(-4). In parallel, pVcNR15-transformed Volvox exhibited growth rates that were 100-fold increased over the pVcNR13-transformed alga. This intron-enhancement of nitA gene expression appears to be associated with post-transcriptional processing and 'channelling' of the message. These data suggest an important role of splicing for gene expression in V. carteri.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ALPHA-TUBULIN GENES; ASEXUAL LIFE-CYCLE; NUCLEAR TRANSFORMATION; CHLAMYDOMONAS-REINHARDTII; REGULATORY ELEMENTS; TRANSGENIC TOBACCO; MAIZE CELLS; CARTERI; RNA; EFFICIENCY; cryptic splice sites; intron-enhancement; gene expression; nitA cDNA; Volvox; transformation |
| Subjects: | 500 Science > 570 Life sciences 500 Science > 580 Botanical sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 Nov 2023 08:37 |
| Last Modified: | 02 Nov 2023 08:37 |
| URI: | https://pred.uni-regensburg.de/id/eprint/51862 |
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