Flow cytometric screening and isolation of Escherichia coli clones which express surface antigens of the oil-degrading microorganism Acinetobacter calcoaceticus RAG-1.
Flow cytometry (FCM) in conjunction with immunocytochemical-labeling was used to analyze and screen a population of Escherichia coli clones containing a genomic library from the oil-degrading microorganism Acinetobacter calcoaceticus RAG-1 surface antigens. Reconstruction experiments using mixed populations indicated that RAG-1 cells could be clearly distinguished at a ratio of one RAG-1 cell to 500 Escherichia coli cells. Using this technique two clones, WM143 and WM191, were isolated and shown by restriction endonuclease cleavage and Southern hybridization to contain plasmids carrying inserts of RAG-1 DNA of 9.4 and 9.8 kb respectively.
Minas
W
W
Department of Microbiology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel.
Sahar
E
E
Gutnick
D
D
D000150:Acinetobacter / Q000235:genetics / Q000276:immunology*; D000942:Antigens, Bacterial / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D005434:Flow Cytometry; D007150:Immunohistochemistry; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015183:Restriction Mapping
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial