Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1985-05-31
1990-09-25
Schwartz, Richard A.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
4351721, 4351723, 935 16, 935 17, C12P 1934
Patent
active
049593129
ABSTRACT:
A method is disclosed for in vitro mutagenesis of a target DNA sequence which generates mutants containing a single randomly-located region in the target sequence with random substitution mutations. The method includes the production of a supply of a template for the target sequence and random primers having differing 3'--OH termini. At least two nucleotides are added randomly to the primers to produce modified random primers, some of which contain at least one mismatch with respect to the template. The modified random primers are employed with the supply of the sequence to polymerize from the 3'--OH terminus of the modified random primer along the template to biologically fix mutations resulting from mismatched bases and produce molecules having a double-stranded region containing a mutant strand. The molecules are transferred into the host organisms to cause at least some of the mutant strands to be replicated in the host organism to produce mutant DNA sequences. These organisms are grown into a population of clones each containing a mutant sequence. One or more clones are selected based on desired characteristics and a mutant sequence is amplified to produce a usable supply of the mutant sequence.
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Schwartz Richard A.
The University of Tennessee Research Corporation
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