Method for transforming bacterium belonging to genus bifidobacterium
AssigneeKabushiki Kaisha Yakult Honsha
InventorsYohei WATANABE, Taeko HARA
Life of this patent
Estimating the term…
Abstract
A method for transforming a bacterium belonging to the genus Bifidobacterium, which includes statically culturing a recipient bacterium belonging to the genus Bifidobacterium in contact with a donor bacterium or linear DNA including DNA to be introduced into the recipient, where a medium for use in the statically culturing is a medium that has pH of 6.0 or higher and lower than 10.0, where a content x (% by mass) of a carbohydrate utilizable by the recipient bacterium belonging to the genus Bifidobacterium in the medium satisfies 0<x≤0.2×(pH)−1.2 when the pH is 6.0 or higher and lower than 8.5 and 0.2×(pH)−1.7<x≤0.2×(pH)−1.2 when the pH is 8.5 or higher and lower than 10.0.
Claims11
A method for transforming a bacterium belonging to the genus Bifidobacterium, the method comprising:
- statically culturing a recipient bacterium belonging to the genus Bifidobacterium in contact with a donor bacterium or linear DNA comprising DNA to be introduced into the recipient,
- wherein a medium for use in the statically culturing is a medium that has pH of from 6.0 to lower than 10.0,
- wherein a content x in mass % of a carbohydrate utilizable by the recipient bacterium belonging to the genus Bifidobacterium in the medium satisfies 0<x≤0.2×(pH)−1.2 when the pH is from 6.0 to lower than 8.5 and 0.2×(pH)−1.7<x≤0.2×(pH)−1.2 when the pH is from 8.5 to lower than 10.0.
A method for manipulating the genome of a bacterium belonging to the genus Bifidobacterium, the method comprising:
- statically culturing a recipient bacterium belonging to the genus Bifidobacterium in contact with donor linear DNA,
- wherein a medium used in the statically culturing is a medium that has pH of from 6.0 to lower than 10.0,
- wherein a content x in mass % of a carbohydrate utilizable by the recipient bacterium belonging to the genus Bifidobacterium in the medium satisfies 0<x≤0.2×(pH)−1.2 when the pH is from 6.0 to lower than 8.5 and 0.2×(pH)−1.7<x≤0.2×(pH)−1.2 when the pH is from 8.5 to lower than 10.0;
- the recipient bacterium belonging to the genus Bifidobacterium is a bacterium belonging to the genus Bifidobacterium having no upp gene;
- and
- wherein the donor linear DNA comprises an upstream homology arm, two genes;
- selection marker gene and upp gene, and a downstream homology arm, wherein the two genes are located between the upstream homology arm and the downstream homology arm, and the donor linear DNA further comprises a 3′-terminal region of the upstream homology arm between the two genes and the downstream homology arm, or a 5′-terminal region of the downstream homology arm between the upstream homology arm and the two genes.
Dependencies are inferred from the claim text.