Mutation oligonucleotide-directed

Natural selection works through the complementary processes of mutation and genetic reassortment by recombination. The oligonucleotide-directed mutagenesis methods used in the foregoing examples do not allow for recombination instead, mutations are combined manually to optimize a protein sequence. Willem Stemmer at Maxygen invented a method of directed evolution that uses both mutation and recombination. This method, called... 

In the Ca-ATPase from sarcoplasmic reticulum, oligonucleotide-directed, site-specific mutagenesis has been applied to identify amino acids involved in Ca binding. Mutation of 30 glutamate and aspartate residues, singly or in groups, in a stalk sector near the transmembrane domain has little effect on Ca " -transport. In contrast mutations to Glu ° , Glu, Asn , Thr , Asp ° or Glu ° resulted in loss... 

Zoller, M. J., and Smith, M., Oligonucleotide-Directed Mutagenesis Using M13-Derived Vectors an Efficient and General Procedure for the Production of Point Mutations in Any Fragment of DNA. Nucleic Acids Res., 1982. 10(20) pp. 6487-6500. 

Cloning of constructs was performed according to standard protocols [Sambrook et al. 1989], The mutations were created in the plasmid pBluescript II KS (+) by oligonucleotide-directed mutagenesis. 

Figure 13-3 The three-dimensional structure of pectate lyase C showing locations of amino acids substituted by oligonucleotide-directed mutation of the cloned gene.

Several point mutations in the S. cerevisiae flavocytochrome coding sequence have been constructed by oligonucleotide-directed site-specific mutagenesis of the cloned gene (143). The enzyme-coding region was transferred to a plasmid designed both for expression of... 

While many diseases have long been known to result from alterations in an individual s DNA, tools for the detection of genetic mutations have only recently become widely available. These techniques rely upon the catalytic efficiency and specificity of enzyme catalysts. For example, the polymerase chain reaction (PCR) relies upon the ability of enzymes to serve as catalytic amplifiers to analyze the DNA present in biologic and forensic samples. In the PCR technique, a thermostable DNA polymerase, directed by appropriate oligonucleotide primers, produces thousands of copies of a sample of DNA that was present initially at levels too low for direct detection. 

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