Method for deletion of a gene from a bacteria
Patent
·
OSTI ID:6061312
This patent describes a method for deletion of a gene from a bacteria. It comprises: constructing a linear DNA fragment wherein the fragment has sequences sufficiently homologous to regions on the cell chromosome flanking the gene to be deleted; to permit base pairing therebetween placing a second gene whose expression results in a rapidly detectable phenotype between the hybridizing sequences in the DNA fragment; introducing the linear DNA fragment into bacterial cells; selecting for a double reciprocal recombination mediated by the RecA gene by culturing the cells containing the linear DNA under conditions wherein the phenotype encoded by the second nucleotide sequence is detected and cells with a double reciprocal recombination express the detectable phenotype; and altering the cell strain phenotype from recA + to reca {minus} immediately after the linear DNA fragment is incorporated into the cell chromosome by introduction of a second linear DNA fragment containing sequences homologous to regions flanking the recA gene.
- Assignee:
- MIT, Cambridge, MA (USA)
- Patent Number(s):
- US 4963487; A
- Application Number:
- PPN: US s 7-096958
- OSTI ID:
- 6061312
- Country of Publication:
- United States
- Language:
- English
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