Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1987-09-14
1990-10-16
Mays, Thomas D.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4351721, 435320, 435849, 536 27, 935 29, 935 73, 935 83, 935 84, C12N 1500, C12N 700, C07H 1512
Patent
active
049634879
ABSTRACT:
Disclosed is a method and linear DNA fragments for use in the deletion of a gene from a bacteria with a single step procedure that is applicable to any essential or nonessential gene which has been cloned. Chromosomal deletions are constructed by transformation of a cell strain with linear DNA fragments containing a locus for resistance to an antibiotic, or any other gene allowing for rapid phenotypic selection, flanked by sequences homologous to closely spaced regions on the cell chromosome on either side of the gene to be deleted, in combination with the immediate subsequent deletion or inactivation of the recA gene. By selecting for a double-crossover event between the homologous sequences, shown by the antibiotic resistance or other detectable phenotype, a chromosome disruption can be selected for which has effectively deleted an entire gene. Inactivation or deletion of the recA gene prevents recombination or incorporation of extrachromosomal elements from occurring, thereby resulting in a bacterial strain which is useful for screening for functional activity or production of genetically engineered proteins in the absence of specific contaminants.
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Massachusetts Institute of Technology
Mays Thomas D.
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