Bifidobacteria CRISPR sequences
The invention relates to CRISPR sequences found in bifidobacteria and their uses.
1. A method for strain typing Bifidobacteria wherein:
the method comprises:
(a) amplifying, using an at least one primer pair, a genomic DNA sequence from a CRISPR locus of a Bifidobacteria strain of interest, wherein said CRISPR locus consists of at least one Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) repeat(s) between two CRISPR spacer(s) wherein said at least one CRISPR repeat consists of:
SEQ ID NO: 2; or
SEQ ID NO: 2 comprising at least one mutation selected from the group consisting of:
substitution of Cytosine for Thymine at position 12,
substitution of Cytosine for Thymine at position 14, and
substitution of Guanine for Adenine at position 36
wherein each primer of the at least one primer pair is complementary to at least a portion of the repeat sequence consisting of SEQ ID NO: 2 or variant thereof, and to a portion of the adjacent spacer, and wherein each primer is selected to create an amplicon comprising at least one spacer sequence of the CRISPR locus; and
(b) detecting the amplicon generated in step (a), whereby said detected amplicon indicates a Bifidobacteria strain type.
2. A method for strain typing Bifidobacteria wherein:
the method comprises:
(a) amplifying, using an at least one primer pair, a genomic DNA sequence from a CRISPR locus of a Bifidobacteria strain of interest, wherein said CRISPR locus consists of at least two CRISPR spacer(s) surrounding one Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) repeat(s), wherein said at least one CRISPR repeat comprises:
SEQ ID NO: 2; or
SEQ ID NO: 2 comprising one or more mutations selected from the group consisting of:
substitution of Cytosine for Thymine at position 12,
substitution of Cytosine for Thymine at position 14, and
substitution of Guanine for Adenine at position 36
wherein each primer of the at least one primer pair is complementary to at least a portion of a different spacer sequence of the CRISPR locus, thereby creating an amplicon comprising at least one repeat sequence located between the two spacer sequences of the CRISPR locus; and
(b) detecting the amplicon generated in step (a), whereby said detected amplicon indicates a Bifidobacteria strain type.
3. The method of claim 1 or 2 , wherein the CRISPR locus is Bala1 comprising SEQ ID NO: 1.
4. The method of claim 1 or 2 , wherein detecting the amplicon includes hybridizing the amplicon to an immobilized probe.
5. The method of claims 1 and 2 , wherein the spacer sequence is selected from the group consisting of SEQ ID NOs: 3 to 24.
6. The method of claim 1 or 2 , further comprising the step of:
(c) detecting the size of the amplicon.