Figure 6.

The mRNA rreB is weakly translated in B. japonicum. (A) Schematic representation of the used constructs cloned between a Paph promoter and a sequence for integration of the plasmids into the chromosome (int).²⁹ The rreB-egfp fusion is preceded either by a standard SD and an ATG start codon (pRJ-sSD-rreB-egfp) or by the full-length 5′-UTR of rreB with the 17c-region and the GTG start codon (pRJ-17c-rreB-egfp). Further modifications of the 5′-UTR of rreB resulting in pRJ-14c-rreB-egfp and pRJ-13c-ATG-rreB-egfp are indicated on the bottom. (B) and (C) Western blot analysis with GFP-specific antibodies of strains containing the indicated constructs (see A). EVC, plasmid pJPaph-MCS was integrated into the chromosome. Schematic representation of of pJH-14c-GTG-egfp is given in Fig. 5A. The bottom panel shows a Coomassie Blue stained SDS-polyacrylamide gel after electrophoresis visualizing the loaded protein amounts. Migration of marker proteins (M) is indicated (in kDa). Arrows indicate the position of the RreB-EGFP band; arrows with asterisks indicate putative degradation products.