Bacillus thuringiensis transformation method
AssigneeNovartis Finance Corporation
InventorsCindy Lou Jellis, Noah D. Beerman, Jean-Christophe Piot
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Abstract
Particularly effective B.t. insecticides are obtained by culturing to the sporulation stage novel transformant obtained on transforming B.t. kurstaki H.D. 562 with DNA expressing in the B.t. certain mutant endotoxin genes. Such transformants and other B.t. transformants generally may be obtained in high transformation frequencies by the electrotransformation of B.t. cells while in a hypertonic state followed by maintaining the cells in a hypertonic medium for a time sufficient to obtain intact cells. Transformation of B.t. tenebroinis and B.t. aizawai is also described.
Claims20
A process for transforming Bacillus thuringiensis (B. thuringiensis) with exogenous DNA comprising establishing a hypertonic aqueous transformation medium containing said host cells and said exogenous DNA to be transformed into said cells, subjecting said transformation medium containing said host cells and said exogenous DNA to a voltage pulsed current of about 1500 to 3000 volts to effect transformation of said host cells by said exogenous DNA, and incubating the thus transformed host cells in a hypertonic aqueous incubation medium for a time sufficient to obtain transformed host cells.
A process for transforming Bacillus thuringiensis with exogenous DNA from B. thuringiensis donor cells comprising establishing a hypertonic aqueous transformation medium containing said host cells and said exogenous DNA to be transformed into said cells, subjecting said transformation medium containing said host cells and said exogenous DNA to a voltage pulsed current between 2200 to 3000 volts to effect transformation of said host cells by said exogenous DNA, and incubating the thus transformed host cells hypertonic aqueous incubation medium for a time sufficient to obtain transformed host cells.
Dependencies are inferred from the claim text.