Expression and secretion of recombinant outer-surface protein A from the Lyme disease agent, Borrelia burgdorferi, in Nicotiana tabacum suspension cells.
The ospA gene of Borrelia burgdorferi codes for an outer membrane lipoprotein, which is a major antigen of the Lyme disease agent. Recombinant OspA vaccines tested so far were expressed in Escherichia coli. In this study, we investigated the expression of a soluble OspA protein in Nicotiana tabacum suspension cells and evaluated the secretion of OspA driven by either its own bacterial signal peptide or a plant signal peptide fused to the amino-terminal cysteine of the mature form. In both cases, the signal peptide was cleaved off and OspA secreted. During secretion, OspA was N-glycosylated. Addition of a C-terminal KDEL sequence led to retention of OspA in the endoplasmic reticulum.
Navarre
Catherine
C
Unité de Biochimie Physiologique, Institut des Sciences de la Vie, Université catholique de Louvain, Croix du Sud 5-15, 1348, Louvain-la-Neuve, Belgium.
Delannoy
Mélanie
M
Lefebvre
Benoit
B
Nader
Joseph
J
Vanham
Delphine
D
Boutry
Marc
M
D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D001428:Bacterial Vaccines / Q000737:chemistry / Q000235:genetics*; D025065:Borrelia burgdorferi / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005821:Genetic Techniques; D005822:Genetic Vectors; D006031:Glycosylation; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics*; D008957:Models, Genetic; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D014026:Nicotiana / Q000378:metabolism* / Q000382:microbiology*; D019076:Transgenes
D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D008074:Lipoproteins; C072045:OspA protein