Identification and characterization of Clostridium difficile promoter element that is functional in Escherichia coli.
The promoter element involved in the expression of a previously characterized cloned clostridial antigen was isolated and characterized. A restriction fragment containing the promoter element of the Clostridium difficile insert was cloned using the promoter probe vector, pGA46. Subclones of the clostridial DNA insert in pGA46 were then analyzed by nucleotide sequencing and by S1 nuclease experiments. The clostridial promoter element exhibits a high degree of homology with typical Escherichia coli promoter elements. This sequence probably represents a unique class of clostridial promoter elements which, given their ability to function in E. coli and C. difficile, can be used in the construction of a shuttle vector capable of gene expression in E. coli and C. difficile.
Dailey
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DC
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46223.
Schloemer
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RH
D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003013:Clostridium / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004720:Endonucleases; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000302:isolation & purification; D015183:Restriction Mapping; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013753:Tetracycline Resistance / Q000235:genetics; D014170:Transformation, Genetic
D012329:RNA, Bacterial; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases