On the forefront of modern scientific innovation, Cloning, Gene Expression, and Protein Purification: Experimental Procedures and Process Rationale effectively...
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Résumé
On the forefront of modern scientific innovation, Cloning, Gene Expression, and Protein Purification: Experimental Procedures and Process Rationale effectively doubles as a laboratory manual for students and a reference book for professional researchers. Designed for advanced undergraduate and beginning graduate students in molecular biology, this unique combination lecture/laboratory resource presents detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein's basic physical properties. This manageable volume includes both theoretical background and practical procedures and is structured around twenty experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on unique chemical properties or substrate affinities. The book describes advanced topics such as the use of antibodies and the techniques developed to transform their structures, as well as combinatorial approaches designed to manipulate the structure and functions of proteins and nucleic acids. Supplemental literature provides a variety of theoretical explanations encouraging a more intuitive understanding of the experimental mechanisms and behaviors of the chemical participants, while also giving students the tools needed to become "capable proactive researchers."
Sommaire
NUCLEIC ACIDS AND CLONING
DNA Isolation
Construction of Recombinant Plasmids
The Polymerase Chain Reaction
Transcription of Genomic DNA and Analysis of the Resulting mRNAs
Transformation and Gene Expression
Analysis of DNA or RNA by Duplex Hybridization: DNA Isolation, Labeling, and Probing
PROTEIN PURIFICATION
Protein Purification
Discontinuous Gel Electrophoresis, Protein Mobilities, and Apparent Size Determination