PEG-mediated plastid transformation: a new system for transient gene expression assays in chloroplasts

✍ Scribed by B. Spörlein; M. Streubel; G. Dahlfeld; P. Westhoff; H. U. Koop

• Book ID: 104672583
• Publisher: Springer
• Year: 1991
• Tongue: English
• Weight: 627 KB
• Volume: 82
• Category: Article
• ISSN: 0040-5752
• DOI: 10.1007/bf00227316

✦ Synopsis

Evidence is presented for the introduction of functional copies of the GUS-reporter gene with plastid regulatory signals into chloroplasts after treatment of Nicotiana plumbaginifolia leaf protoplasts with PEG. GUS-activity is found in cells derived from protoplasts treated with PEG in the presence of plasmids harbouring the GUS-gene under the control of plastid promoter and terminator signals (plastid-specific reporter gene constructions). The activity is maintained after chloroplast isolation and incubation with the protease thermolysin under conditions sufficient to completely remove the much higher transient nuclear/cytoplasmic expression of a GUS-gene carrying the CaMV 35S-promoter. Likewise, GUS-activity derived from a plasmid coding for the nuclear/cytoplasmic expression of the reporter gene with a plastid transit presequence is also maintained after these procedures. These results indicate that PEG-treatment is a suitable protocol by which to introduce DNA into chloroplasts for the study of transient gene expression.