Development of an In Vitro Chloroplast Splicing System: Sequences Required for Correct pre-mRNA Splicing.

ABSTRACT: Chloroplast genomes in land plants include approximately 20 intron-containing genes. Most of the introns are similar to the group II introns found in fungi, algae and some bacteria, but no self-splicing has been reported. To analyze splicing reactions in chloroplasts, we developed a tobacco (Nicotiana tabacum) chloroplast-based in vitro system. We optimized the splicing reaction using atpF precursor messenger RNA (pre-mRNA). Our system requires a high ATP concentration, whereas ATP is not necessary for self-splicing group II introns. Self-splicing group II introns possess two exon-binding sites (EBS1 and 2) complementary to two intron-binding sites (IBS1 and 2) in the 3' end of 5' exons, which are involved in 5' splice-site selection. Using our in vitro system and atpF pre-mRNA, we analyze

SUBMITTER: Inaba-Hasegawa K 

PROVIDER: S-EPMC8579278 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature