Identification of an endoflagellar associated protein in Borrelia burgdorferi.
DNA of Borrelia burgdorferi was cleaved by the endonuclease EcoRI and ligated with the bacteriophage expression vector lambda gt11. After infection of the Escherichia coli strain Y1089, the plaques of recombinant phages were screened with a B. burgdorferi antiserum (human) for fusion proteins containing borrelia antigen.s A positive clone produced a hybrid protein (p200) of c. 200 Kda. The corresponding native borrelia protein (p97) was identified as having an Mr of 97 Kda. To localise protein p97 in the B. burgdorferi cell, immunoelectronmicroscopy and a Western blot of isolated flagella were used. Antibodies directed against proteins p200 and p97 recognised epitopes associated with the flagella.
Eiffert
H
H
Department of Medical Microbiology, University of Göttingen, Germany.
Schlott
T
T
Hoppert
M
M
Lotter
H
H
Thomssen
R
R
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D015153:Blotting, Western; D015748:Borrelia burgdorferi Group / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D004274:DNA, Recombinant / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI; D004586:Electrophoresis; D000939:Epitopes / Q000032:analysis; D005407:Flagella / Q000737:chemistry*; D005822:Genetic Vectors; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013987:Ticks / Q000382:microbiology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D015246:Deoxyribonuclease EcoRI