Jakobiak, Thomas and Mages, Wolfgang and Scharf, Birgit and Babinger, Patrick and Stark, Klaus and Schmitt, Ruediger (2004) The bacterial paromomycin resistance gene, aphH, as a dominant selectable marker in Volvox carteri. PROTIST, 155 (4). pp. 381-393. ISSN 1434-4610,
Full text not available from this repository. (Request a copy)Abstract
The aminoglycoside antibiotic paromomycin that is highly toxic to the green alga Volvox carteri is efficiently inactivated by aminoglycoside 3'-phosphotransferase from Streptomyces rimosus. Therefore, we made constructs in which the bacterial aphH gene encoding this enzyme was combined with Volvox cis-regulatory elements in an attempt to develop a new dominant selectable marker - paromomycin resistance (Pm-R) - for use in Volvox nuclear transformation. The construct that provided the most efficient transformation was one in which aphH was placed between a chimeric promoter that was generated by fusing the Volvox hsp70 and rbcS3 promoters and the 3' UTR of the Volvox rbcS3 gene. When this plasmid was used in combination with a high-impact biolistic device, the frequency of stable Pm-R transformants ranged about 15 per 106 target cells. Due to rapid and sharp selection, Pm-R transformants were readily isolated after six days, which is half the time required for previously used markers. Co-transformation of an unselected marker ranged about 30%. The chimeric aphH gene was stably integrated into the Volvox genome, frequently as tandem multiple copies, and was expressed at a level that made selection of Pm-R transformants simple and unambiguous. This makes the engineered bacterial aphH gene an efficient dominant selection marker for the transformation and co-transformation of a broad range of V carteri strains without the recurring need for using auxotrophic recipient strains.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GREEN-ALGA VOLVOX; CHLAMYDOMONAS-REINHARDTII; ANTIBIOTIC-RESISTANCE; NITRATE REDUCTASE; EXPRESSION; PROTEIN; PLAYS; DIFFERENTIATION; TRANSFORMATION; PATTERNS; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 21 Jun 2021 13:03 |
| Last Modified: | 21 Jun 2021 13:03 |
| URI: | https://pred.uni-regensburg.de/id/eprint/36935 |
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