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. 2023 Feb 21;13(3):315. doi: 10.3390/metabo13030315

Figure 1.

Figure 1

Southern blot analysis of transplastomic lines. (A) Schematic representation of the insertion locus. Numbers refer to positions on the tobacco chloroplast genome, accession Z00044.2 (drawn to scale). (B) Schematic representation of the LTB::L1-HPV16B transgene cassette inserted into the neutral EcoRV site shown in (A). The ndhB sRNA is shown in green. P16S = promoter of the chloroplast 16S rRNA gene. 3′rbcL = the 3′-UTR of the rbcL mRNA from Chlamydomonas reinhardtii. Arrows point in the direction of transcription of the respective genes. (C) same as in (B), but for the L1-HPV18.2 transgene cassette. (D) same as in (B), but for the RV5 control construct. (E) Sequence excerpts from L1-HPV18.2 and LTB::L1-HPV16B transgene cassettes. (F) RFLP analysis of transplastomic lines, where 5.0 µg of cellular DNA was fragmented by SalI. Hybridization was performed with a radiolabeled probe derived from the petG and trnW genes (bar in (A), probe position 68,595–68,872 of Z00044.2). Numbers on the right side of the blots indicate the expected size for restriction fragments: 8670 bp = LTB::L1-HPV16B fragment; 8307 bp = L1-HPV18.2 fragment; 6380 bp = RV5 fragment; 5471 bp = wt fragment. (G) Plants were grown on sucrose-containing MS medium for three weeks (upper row) or germinated on soil and grown for two weeks (lower row).