directed mutagenesis and protein engineering

Examples of Properties Altered for Improved Therapeutic or Industrial Applications. Michaelis constant (Km), Vmax, catalytic rate constant (kcat)Thermal tolerance, pH stabilityReactivity in nonaqueous solventsEliminate cofactor requirementAlter substrate binding site/specificityIncrease protease resistanceAlter allosteric regulation.

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Site-directed Mutagenesis

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Mutagenesis Methods

Mutagenesis Methods. Lily Peterson April 5 th , 2010. Overview: Types of Mutagenesis. PCR Based Methods Site-directed mutagenesis Mismatched mutagenesis 5’ add on mutagenesis Cassette Mutagenesis Insertional Mutagenesis Trasposon mutagenesis In vivo Mutagenesis Direct Mutagenesis.

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Mutations, Mutagenesis, and Repair

Mutations, Mutagenesis, and Repair. Chapter 10 . The Problem. DNA extremely long, fragile Subject to both physical and chemical damage Consequences could be lethal for organism or offspring. }. Frameshift mutations. Mutation. A heritable change in the base sequence of DNA

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Directed Mutagenesis and Protein Engineering

Directed Mutagenesis and Protein Engineering. Mutagenesis. Mutagenesis -> change in DNA sequence -> Point mutations or large modifications Point mutations (directed mutagenesis): Substitution: change of one nucleotide (i.e. A-> C) Insertion: gaining one additional nucleotide

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Genomes and mutagenesis

Genomes and mutagenesis. Figure 9.15. The Ames test relies on a mutant bacterial strain that is defective in hisG. - Cannot grow on a medium lacking histidine. Identifying Mutagens. - A mutagen-containing disk is placed on an agar plate with the mutant.

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MUTAGENS AND MUTAGENESIS

MUTAGENS AND MUTAGENESIS. Mutagens. Naturally occurring mutations are referred as spontaneous mutations and are thought to arise through chance errors in chromosomal division or DNA replication. Environmental agents which cause mutations are known as mutagens

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Site-directed mutagenesis

Site-directed mutagenesis. Mutation is a random process The rate of mutation can be increased with radiation or mutagenic chemicals Individuals mutant at certain loci can be selected for or screened for, but the exact location and nature of the mutation is unknown.

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Genome-Scale Mutagenesis

Introduction Model systems Yeast Mouse Implications for science. Genome-Scale Mutagenesis. what is a gene? genes to function how do you study this?. Genotype - Phenotype. Reverse Genetics - Forward Genetics. Reverse:. Genotype Single gene locus Hemoglobin CFTR Rb

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Mouse mutagenesis

Mouse mutagenesis. Roberta Rivi, MD Laboratory of Molecular Embryology. Mutagenesis?. Goal: determination of the function of every gene ( functional genomics ) Gene-driven also called reverse genetics Phenotype-driven also called forward genetics (classical approach).

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Protein Function Analysis using Computational Mutagenesis

Protein Function Analysis using Computational Mutagenesis. Iosif Vaisman Laboratory for Structural Bioinformatics proteins.gmu.edu. CASB workshop, 9/23/10. Department of Bioinformatics and Computational Biology. Dealunay simplices classification. Protein representation (Crambin).

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Supplementary figure S6 Attempts to improve solubility of DXS by protein mutagenesis

p > 0.1. p > 0.1. Supplementary figure S6 Attempts to improve solubility of DXS by protein mutagenesis

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Site Directed Mutagenesis

Site Directed Mutagenesis. What Is a Mutation?. What Is a Mutation?. What Is a Mutation?.

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MUTAGENESIS & MUTATIONS

MUTAGENESIS & MUTATIONS. http://biolmolgen.slam.katowice.pl. http://www.elearning.sum.edu.pl/www/professorpage.php. Aleksander L. Sieroń Katedra Biologii Molekularnej i Genetyki , Ś U M Katowice. MUTAGEN.

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Protein Engineering : Site-Directed Mutagenesis dan Directed Evolution

Protein Engineering : Site-Directed Mutagenesis dan Directed Evolution. Dessy Natalia Departemen Kimia dan KPP Bioteknologi Institut Teknologi Bandung. Keanekaragaman Protein. Keanekaragaman hayati Rekayasa Protein In silico. Rational Protein Design.

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QTL analysis / Mutagenesis

Genetic basis of atherosclerotic organ damage: From animal genetics to human cohorts. QTL analysis / Mutagenesis. atherosclerosis renal damage HDL cholesterol PLTP activity. candidate genes. functional studies. functional studies. Human populations. Principle of QTL analysis (1).

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Mutagenesis and Genetic Screens

Mutagenesis and Genetic Screens. General pathway for mutational dissection of a biological process “Forward Genetics”. Fig. 12-39. General pathway for mutational dissection of a biological process “Forward Genetics”. Fig. 12-39. Classification of Mutant Type.

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Site Directed Mutagenesis of Protein PurE

Site Directed Mutagenesis of Protein PurE. Megan Silas From the University of Illinois at Urbana Champaign In Dr. Fung’s Lab in the Department of Chemistry. Outline. Project Overview Bacillus anthracis Purines PurE Experimental Procedures and Results Primer Design

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site directed mutagenesis

Random Mutagenesis Creative Biogene uses the error-prone PCR technique to introduce mutations into a gene, result in amino acid substitutions in proteins. The mutated PCR products are cloned into the specific vector of your choice. Creative Biogene can generate 1 to 4 random amino acid substitutions per 1,000bp. And the randomized portion of the gene can cover the complete open reading frame or your desired regions of the gene. Creative Biogene can offer the most affordable and fastest random mutagenesis service for your scientific research.

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Directed Mutagenesis and Protein Engineering

Directed Mutagenesis and Protein Engineering. Mutagenesis. Mutagenesis -> change in DNA sequence -> Point mutations or large modifications Point mutations (directed mutagenesis): Substitution: change of one nucleotide (i.e. A-> C) Insertion: gaining one additional nucleotide

378 views • 31 slides

Mouse mutagenesis

Mouse mutagenesis. Roberta Rivi, MD Laboratory of Molecular Embryology. Mutagenesis?. Goal: determination of the function of every gene ( functional genomics ) Gene-driven also called reverse genetics Phenotype-driven also called forward genetics (classical approach).

252 views • 24 slides

Site Directed Mutagenesis of Protein PurE

Site Directed Mutagenesis of Protein PurE. Megan Silas From the University of Illinois at Urbana Champaign In Dr. Fung’s Lab in the Department of Chemistry. Outline. Project Overview Bacillus anthracis Purines PurE Experimental Procedures and Results Primer Design

357 views • 26 slides