Production of Site Specific Nucleases,CAP Protein,Iodoacetamide,Phenanthroline,DNA Cleaving,Nucleases,Staphylococcus,Hydrolyses,DNA or RNA

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Production of Site Specific Nucleases (e.g. Restriction Enzymes)
The DNA recognition and binding properties of proteins can be combined using chemical cleavage agents. Cys178 of E. coli CAP protein; has been modified using '5-iodoacetamide -1, 10phenanthroline' yielding a DNA-cleaving agent that recognized and cleaved DNA at the centre of the recognition site (22 bp) for CAP. This may give restriction enzymes recognizing upto 20 bases instead of 6 or 8 bases and may, therefore, be useful for isolating long DNA fragments needed for sequencing and mapping.

Nucleases may also be produced by fusion of non-specific phosphodiesterases to oligonucleotides of defined sequence. For a nuclease from Staphylococcus modified by this approach, it was shown that oligonucleotide component of fused product pairs with its complementary sequence and the hybrid enzyme hydrolyses single stranded DNA or RNA adjacent to the oligonucleotide binding site. This approach thus can also be used for developing artificial restriction enzymes.

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Methods for Protein Engineering Site Directed Mutagenesis in Vivo Gene Modification Through Oligonucleotide Synthesis Breeding a Better Catalyst Using Random Events Artificial Random Mutagenesis Random DNA Shuffing Using Recombination Combination of Structure Based and Envolutionary Approaches Multienzyme Systems bi and Polyfunctional Enzymes by Gene Fusion Chemical Modification of Enzymes Production of Site Specific Nucleases Production of Artifical Semi Synthetic Oxido Reductases Flavo Enzyme Hybrid Enzymes Combinatorial Chemistry Organic Synthesis Libraries De Novo Design of Catalysts Some Early Achievements of Protein Engineering Possible Candidates for Future Protein Engineering Improving Enzymes by Using Organic Solvents