Native promoter-plasmid vector system for heterologous cholesterol oxidase synthesis in Streptococcus thermophilus.
The cholesterol oxidase gene (choA) of a streptomycete was used as a model for studying heterologous gene expression in Streptococcus thermophilus, an essential bacterium in dairy food fermentations. The vectors pER82 and pER82P were developed from the 2.2-kb indigenous plasmid (pER8) of S. thermophilus ST108, and sP1, a 51-bp synthetic promoter patterned after a chromosomal sequence of S. thermophilus. The presence of sP1 promoter in pER82PbCOb with the choA insert aligned with the cat gene was essential for the intracellular production of cholesterol oxidase. The pER82PbCOb was apparently stable in S. thermophilus with no detectable evidence of deletion mutational events.
Somkuti
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U.S. Department of Agriculture, ARS, Eastern Regional Research Center, Philadelphia, Pennsylvania 19118, USA.
Solaiman
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Steinberg
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D015139:Blotting, Southern; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D002789:Cholesterol Oxidase / Q000096:biosynthesis* / Q000235:genetics; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D013291:Streptococcus / Q000201:enzymology / Q000235:genetics*; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D002789:Cholesterol Oxidase; D015500:Chloramphenicol O-Acetyltransferase