Biochemical procedure for production of small deletions in simian virus 40 DNA.
A simple biochemical procedure for producing small deletions (15 to 50 base pairs) at virtually any location in simian virus 40 DNA has been developed. The steps involved are: cleavage of the closed-circular DNA to produce a linear structure followed by 5'-exonuclease digestion to expose a short single-stranded segment at each 3' end of the molecule. Mutants containing deletions at the site of the cleavage are obtained by infecting permissive monkey kidney cells with the exonuclease-treated DNA in the presence or absence of a helper DNA (depending upon whether or not the site of cleavage and therefore the deletion occurred in a gene required for vegetative multiplication). In this paper viable mutants with deletions at the HpaII endonuclease cleavage site (0.735 map position) and defective trans-complementable mutants with deletions at the EcoRI endonuclease cleavage site (0/1.0 map position) were isolated.
D003090:Coliphages / Q000201:enzymology; D004270:DNA, Circular / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D003851:Deoxyribonucleases; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases; D009154:Mutation; D013539:Simian virus 40 / Q000378:metabolism*; D013045:Species Specificity; D010948:Viral Plaque Assay
D004270:DNA, Circular; D004279:DNA, Viral; D003851:Deoxyribonucleases; D004720:Endonucleases