RRC ID 47412
著者 Zienkiewicz M, Krupnik T, Drożak A, Golke A, Romanowska E.
タイトル Transformation of the Cyanidioschyzon merolae chloroplast genome: prospects for understanding chloroplast function in extreme environments.
ジャーナル Plant Mol Biol
Abstract KEY MESSAGE:We have successfully transformed an exthemophilic red alga with the chloramphenicol acetyltransferase gene, rendering this organism insensitive to its toxicity. Our work paves the way to further work with this new modelorganism. Here we report the first successful attempt to achieve a stable, under selectable pressure, chloroplast transformation in Cyanidioschizon merolae-an extremophilic red alga of increasing importance as a new model organism. The following protocol takes advantage of a double homologous recombination phenomenon in the chloroplast, allowing to introduce an exogenous, selectable gene. For that purpose, we decided to use chloramphenicol acetyltransferase (CAT), as chloroplasts are particularly vulnerable to chloramphenicol lethal effects (Zienkiewicz et al. in Protoplasma, 2015, doi: 10.1007/s00709-015-0936-9 ). We adjusted two methods of DNA delivery: the PEG-mediated delivery and the biolistic bombardment based delivery, either of these methods work sufficiently with noticeable preference to the former. Application of a codon-optimized sequence of the cat gene and a single colony selection yielded C. merolae strains, capable of resisting up to 400 µg/mL of chloramphenicol. Our method opens new possibilities in production of site-directed mutants, recombinant proteins and exogenous protein overexpression in C. merolae-a new model organism.
巻・号 93(1-2)
ページ 171-183
公開日 2017-1-1
DOI 10.1007/s11103-016-0554-8
PII 10.1007/s11103-016-0554-8
PMID 27796719
PMC PMC5243890
MeSH Biolistics Chloramphenicol O-Acetyltransferase / genetics Chloroplasts / genetics* Chloroplasts / physiology Genome, Chloroplast* Homologous Recombination Plants, Genetically Modified Rhodophyta / genetics* Transformation, Genetic
IF 3.302
引用数 13
WOS 分野 PLANT SCIENCES BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
藻類 NIES-1332