Heterologous expression of genes encoding bacterial light-harvesting complex II in Rhodobacter capsulatus and Rhodovulum sulfidophilum.
In the present work we report the high-level expression of foreign genes encoding the light-harvesting (LHII) membrane-spanning polypeptides in photosynthetic bacteria. To do this we first constructed three deletion strains of Rhodovulum (Rhv.) sulfidophilum in which all or part of the puc operon, encoding the peripheral light-harvesting proteins, is missing. To investigate the heterologous expression of the light-harvesting polypeptides from Rb. capsulatus in Rhv. sulfidophilum and vice versa we have reintroduced functional foreign LH genes into these and equivalent strains of Rhodobacter (Rb.) capsulatus. In some cases very high levels of expression were obtained (85%) of those observed in the wild type), while in other cases much lower expression was observed; possible reasons for these differences are discussed. The heterologously expressed proteins were shown to contain normal pigment-binding sites and to be normally and functionally integrated within the host photosynthetic apparatus. The results indicate that heterologous proteins are able to assemble properly and enter into the same protein-protein interactions as their analogs originally present in the host strain.
Katsiou
E
E
Institut für Biologie II, Albert-Ludwigs-Universität, Freiburg, Germany.
Sturgis
J N
JN
Robert
B
B
Tadros
M H
MH
D001426:Bacterial Proteins; D015152:Blotting, Northern; D015139:Blotting, Southern; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins / Q000737:chemistry; D016296:Mutagenesis / Q000235:genetics; D009693:Nucleic Acid Hybridization; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids / Q000737:chemistry; D012329:RNA, Bacterial / Q000737:chemistry; D015183:Restriction Mapping; D016354:Rhodobacter capsulatus / Q000737:chemistry / Q000235:genetics*; D012244:Rhodospirillaceae / Q000737:chemistry / Q000235:genetics*; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013059:Spectrum Analysis, Raman
C081521:B800-850 light-harvesting complex, bacteria; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D045342:Light-Harvesting Protein Complexes; D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D012329:RNA, Bacterial