Mapping of transcribed regions of Euglena gracilis chloroplast DNA

Abstract

RNA transcription from defined regions of the E. gracilis chloroplast genome was characterized by hybridization of total cell RNA to 3H-labeled chloroplast DNA restriction endonuclease fragments. Chloroplast DNA was digested into 5 fragments of 53, 35, 25, 10 and 6.9 kilobase pairs (kbp) with Pst1. The 53 kbp DNA was also subfractionated by BamHI digestion. The extent of transcription of the Pst1 fragments was found to be 30, 17, 15, 2.2 and 2.3 kb of RNA, respectively. The total amount of RNA transcription of 67 kb represents 26% of the double-strand information content of the genome. Transcribed regions are dispersed throughout the DNA. The RNA transcripts are present in 2 major abundance classes in the cell. High abundance transcripts of approximately 106 copies/cell were mapped in the rRNA gene region of the 53 kbp fragment and in the 35 kbp fragment. Low abundance transcripts of approximately 1000-4000 copies/cell were mapped in all 5 Pst fragments.