Construction and initial analysis of a representative lambda ZAPII expression library of the intracellular rickettsia Cowdria ruminantium: cloning of map1 and three other Cowdria genes.
The causative agent of heartwater, the rickettsia Cowdria ruminantium, is very poorly understood at the molecular level owing to a profound lack of suitable tools. We have developed an immunoaffinity chromatographic method to purify C. ruminantium from host cell components and the purified rickettsial cells have been used to prepare substantially pure Cowdria DNA. This DNA has been used to construct what we believe to be the first fully representative C. ruminantium expression library. A clone containing the complete Cowdria map1 gene has been isolated and sequenced. This gene has been expressed in E. coli cells from the native Cowdria promoter, suggesting that the mechanisms for gene transcription and translation are similar between these two organisms. Parts of three other Cowdria genes have also been isolated and sequenced.
Brayton
K A
KA
Onderstepoort Veterinary Institute, South Africa.
Fehrsen
J
J
de Villiers
E P
EP
van Kleef
M
M
Allsopp
B A
BA
D000818:Animals; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics*; D010582:Bacteriophage lambda; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D016995:Ehrlichia ruminantium / Q000235:genetics* / Q000302:isolation & purification; D015723:Gene Library; D006357:Heartwater Disease / Q000382:microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D012675:Senegal
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D011994:Recombinant Proteins; C086402:major antigenic protein 1, Cowdria ruminantium