The atpF group-II intron-containing gene from spinach chloroplasts is not spliced in transgenic Chlamydomonas chloroplasts.
In order to determine whether the group-II trans-splicing machinery of the chloroplast of Chlamydomonas reinhardtii can splice a heterologous group-II cis intron, the atpF gene of spinach was transferred into the chloroplast genome of C. reinhardtii using the atpX expression vector. The atpF gene contains a group-II intron which, like other higher plant chloroplast introns, does not self-splice in vitro. The chimeric transgene was expressed at high levels, based on the accumulation of the precursor; however, spliced products could not be detected by Northern blotting, or by RT-PCR coupled with Southern-blot hybridization of the amplified products with an exon-junction probe. These results indicate that the spinach atpF intron is not spliced in transgenic C. reinhardtii chloroplasts. Thus, splicing of chloroplast introns mediated by cellular factors may be species-specific; alternately, the group-II splicing machinery of C. reinhardtii is specific for trans spliced introns.
Deshpande
N N
NN
Department of Botany, University of Texas at Austin 78713, USA.
Hollingsworth
M
M
Herrin
D L
DL
D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism; D007438:Introns; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010940:Plant Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012326:RNA Splicing; D018724:Spinacia oleracea / Q000235:genetics*; D014170:Transformation, Genetic; D019076:Transgenes
D009838:Oligodeoxyribonucleotides; D010940:Plant Proteins; D006180:Proton-Translocating ATPases