Strains of Lactobacillus Rhamnosus GG Capable of Growth on Lactose and the Uses Thereof
OwnerMatthew Ian Heidtman, Massimo Merighi +1inventors
InventorsMatthew Ian Heidtman, Massimo Merighi, John M. McCoy
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Abstract
The invention provides compositions and methods utilizing an improved strain of bacteria, a Lac+ Lactobacillus rhamnosus strain of bacteria.
Claims20
A Lactobacillus rhamnosus bacterial strain comprising at least 99% identity to the nucleotide sequence of strain ATCC 53103 and further comprising a gene encoding a functional LacG protein and a gene encoding a functional LacT protein or a lacTEGF operon lacking a functional transcriptional terminator within 100 nucleotide base pairs from the initiating codon of lacT.
A Lactobacillus rhamnosus bacterial strain comprising at least 99% of the nucleotide sequence of strain ATCC 53103 and further comprising one or more of the following:
- a. at least one mutation that suppresses the LacT defect found in the strain ATCC 53103;
- b. at least one mutation in the transcriptional terminator regulating the lacTEGF operon, wherein the at least one mutation inactivates the transcriptional terminator;
- c. the nucleotide sequence encoding a functional LacT or the amino acid SEQ ID NO:
- 11;
- and
- d. the nucleotide sequence encoding a functional LacG or the amino acid sequence of SEQ ID NOS:
- 6 or 8.
A method of identifying a bacteria that can digest lactose comprising:
- a. transforming a Lactobacillus rhamnosus with a first plasmid comprising functional or wild-type LacT and a selective marker;
- b. selecting for at least one Lac+ colony by growing the transformants of step (a) on plates containing lactose and the selective marker,
- c. passaging the Lac+ colony of step (b) at least two times in the absence of lactose and the selective marker, such that the Lac+ colonies no longer contain the first plasmid;
- d. growing the Lac+ colony of step (c) on plates containing lactose, thereby identifying a bacteria that can digest lactose.
Dependencies are inferred from the claim text.