Skip Navigation

This Article
Right arrow Print PDF (2848K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Bock, R.
Right arrow Articles by Maliga, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bock, R.
Right arrow Articles by Maliga, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1995, Vol. 23, No. 13 2544-2547
© 1995

METHODS

Correct splicing of a group II intron from a chimeric reporter gene transcript in tobacco plastids

Ralph Bock+ and Pal Maliga*

Waksman Institute, Rutgers, The State University of New Jersey PO Box 759, Old Hoes Lane, Piscataway, NJ 08855–0759, USA

*To whom correspondence should be addressed

Received February 8, 1995. Accepted May 10, 1995.

A in vivo test system was developed to study group II Intron splicing in higher plant chloroplasts. The chimeric reporter gene uldA* was constructed by translatlonal fusion of an intron-containlng segment of the plastld atpF gene with the coding region of a plastld uldA reporter gene. The chimeric uldA* gene was inserted into the tobacco plastid genome by the biollstlc transformation procedure using a plastid targeting vector. Correct intron excision was confirmed by Northern blot analysis, by sequencing amplified DNAs and by accumulation of the encoded fi-glucuronldase (GUS), the expression of which was dependent on intron removal. Removal of the intron from the uldA*mRNA is less efficient (<50%) than from the atpF mRNA (>90%). The efficiency of atpF mRNA splicing is not affected in the plastid transformants Indicating that inefficient splicing of the highlyexpressed uldA* mRNA is not due to depletion of factors) required for the atpF intron removal. A derivative of uldA*, with a stop codon introduced into the loop of domain VI, was also tested. The mutations did not affect the splicing efficiency. The chimeric uldA* splicing system will facilitate the study of structural and sequence requirements for group II Intron splicing in plastids of higher plants.

+Present address: Institut für Biologie III, Universität Freiburg, Schänzlestraße 1, D-79104 Freiburg, Germany


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
J. Vogel, T. Borner, and W. R. Hess
Comparative analysis of splicing of the complete set of chloroplast group II introns in three higher plant mutants
Nucleic Acids Res., October 1, 1999; 27(19): 3866 - 3874.
[Abstract] [Full Text] [PDF]

Home page
 JCBHome page
S. Ruf, H. Kossel, and R. Bock
Targeted Inactivation of a Tobacco Intron-containing Open Reading Frame Reveals a Novel Chloroplast-encoded Photosystem I-related Gene
J. Cell Biol., October 6, 1997; 139(1): 95 - 102.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.