Biochemistry and molecular biology international, 1995-08, Vol.36 (5), p.1025-1035
1995
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- Autor(en) / Beteiligte
- Titel
- Insertion mutagenesis of Chlamydomonas reinhardtii by electroporation and heterologous DNA
- Ist Teil von
- Biochemistry and molecular biology international, 1995-08, Vol.36 (5), p.1025-1035
- Ort / Verlag
- England
- Erscheinungsjahr
- 1995
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- We introduced plasmid pCrmVCAT containing a chloramphenicol acetyltransferase (CAT) gene, flanked by the cauliflower mosaic virus 35S promoter and nopaline synthase polyadenylation sequences, into Chlamydomonas reinhardtii by electroporation; chloramphenicol (CAM) resistance was used for selection. Several mutants with aberrant response to cadmium (Cd) toxicity were obtained by screening CAM resistant transformants. Southern blot analysis showed random integration of pCmVCAT sequence into the nuclear genome. Expression of CAT gene was confirmed by the detection of CAT gene transcript in Northern blot analysis and the detection of CAT enzyme by ELISA assay. This study demonstrated the feasibility of transforming Chlamydomonas reinhardtii with heterologous DNA by electroporation, and the expression of heterologous gene, in this alga.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1039-9712Titel-ID: cdi_pubmed_primary_7580998
- Format
- –
- Schlagworte
- Animals, Cadmium - toxicity, Cadmium Chloride, Cauliflower mosaic virus, Chlamydomonas reinhardtii, Chlamydomonas reinhardtii - drug effects, Chlamydomonas reinhardtii - genetics, Chloramphenicol - pharmacology, chloramphenicol acetyltransferase, Chloramphenicol O-Acetyltransferase - genetics, Chloramphenicol O-Acetyltransferase - metabolism, Chloramphenicol Resistance, Chlorides - toxicity, DNA, Protozoan - genetics, DNA, Protozoan - isolation & purification, DNA, Recombinant - genetics, Electroporation, genes, genetic transformation, Genetic Vectors - genetics, Mutagenesis, Insertional, plasmids, Plasmids - genetics, Plasmids - metabolism, RNA, Messenger - analysis, RNA, Protozoan - analysis, stability, Transfection - methods