Biotechnology

Biotechnology
Author: David Wilson Burden
Publisher:
Total Pages: 317
Release: 1995-01-01
Genre: Molecular cloning
ISBN: 9783764338435

Recombinant Proteins from Plants

Recombinant Proteins from Plants
Author: Charles Cunningham
Publisher: Springer Science & Business Media
Total Pages: 309
Release: 2008-02-05
Genre: Science
ISBN: 1603272607

Recombinant Proteins from Plants is one of the most exciting and fastest developing areas in biology. The latest molecular techniques are being applied to the exploitation of plants as novel expression systems for the p- duction and overproduction of heterologous and native proteins. Transgenic plant technology is currently used in three broad areas: the expression of - combinant proteins to improve crop quality by increasing disease/pest res- tance or increasing tolerance to stress, optimizing plant productivity and yield by the genetic manipulation of metabolic pathways, and the large-scale co- effective production of recombinant proteins for use as specialist industrial or therapeutic biomolecules. The intention of Recombinant Proteins from Plants is to provide c- prehensive and detailed protocols covering all the latest molecular approaches. Because the production oftransgenic plants has become routine in many la- ratories, coverage is also given to some of the more "classical" approaches to the separation, analysis, and characterization of recombinant proteins. The book also includes areas of research that we believe will become increasingly important in the near future: efficient transformation of monocots with Agrobacterium optimizing the stability of recombinant proteins, and a section highlighting the immunotherapeutic potential of plant-expressed proteins.

PCR

PCR
Author: Lucília Domingues
Publisher: Springer Nature
Total Pages: 255
Release: 2023-09-23
Genre: Science
ISBN: 1071633589

This second volume focuses on PCR methods and PCR application specificities to the biotechnology and bioengineering field. New and updated chapters detail real-time PCR protocols, synthetic biology applications, pathogen detection, microfluidics, digital, multiplex detection recent advances. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, PCR: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

Protein Engineering Protocols

Protein Engineering Protocols
Author: Kristian Müller
Publisher: Springer Science & Business Media
Total Pages: 318
Release: 2007-10-26
Genre: Science
ISBN: 1597451878

Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general.

A Practical Guide to Protein Engineering

A Practical Guide to Protein Engineering
Author: Tuck Seng Wong
Publisher: Springer Nature
Total Pages: 213
Release: 2020-10-29
Genre: Science
ISBN: 3030568989

This textbook introduces readers in an accessible and engaging way to the nuts and bolts of protein expression and engineering. Various case studies illustrate each step from the early sequence searches in online databases over plasmid design and molecular cloning techniques to protein purification and characterization. Furthermore, readers are provided with practical tips to successfully pursue a career as a protein engineer. With protein engineering being a fundamental technique in almost all molecular biology labs, the book targets advanced undergraduates and graduate students working in molecular biology, biotechnology and related scientific fields.

Calculations for Molecular Biology and Biotechnology

Calculations for Molecular Biology and Biotechnology
Author: Frank H. Stephenson
Publisher: Academic Press
Total Pages: 520
Release: 2010-07-30
Genre: Science
ISBN: 012375691X

Calculations for Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory, Second Edition, provides an introduction to the myriad of laboratory calculations used in molecular biology and biotechnology. The book begins by discussing the use of scientific notation and metric prefixes, which require the use of exponents and an understanding of significant digits. It explains the mathematics involved in making solutions; the characteristics of cell growth; the multiplicity of infection; and the quantification of nucleic acids. It includes chapters that deal with the mathematics involved in the use of radioisotopes in nucleic acid research; the synthesis of oligonucleotides; the polymerase chain reaction (PCR) method; and the development of recombinant DNA technology. Protein quantification and the assessment of protein activity are also discussed, along with the centrifugation method and applications of PCR in forensics and paternity testing. Topics range from basic scientific notations to complex subjects like nucleic acid chemistry and recombinant DNA technology Each chapter includes a brief explanation of the concept and covers necessary definitions, theory and rationale for each type of calculation Recent applications of the procedures and computations in clinical, academic, industrial and basic research laboratories are cited throughout the text New to this Edition: Updated and increased coverage of real time PCR and the mathematics used to measure gene expression More sample problems in every chapter for readers to practice concepts

Production Technology of Recombinant Therapeutic Proteins

Production Technology of Recombinant Therapeutic Proteins
Author: Chiranjib Chakraborty
Publisher: Daya Books
Total Pages: 290
Release: 2004
Genre: Biotechnology
ISBN: 9788176221047

An Increasing Number Of Recombinant Therapeutic Proteins Are Currently Being Developed, Tested In Clinical Trials And Marketed For Used. Most Of The Recombinant Therapeutic Proteins Are Being Successfully Produced Into Escherichia Coli And Pichia Pastoris Expression System. These Two Expression Systems Are Very Much Efficient And Cost Effective. This Book Takes A Close Look Of These Two Expression Systems And Fermentation Conditions, Purification Strategies Of Different Recombinant Proteins. This Book Also Discusses The Market Size And Cost Analysis For The Production Of Different Therapeutic Proteins And Some General Experimental Protocols For Production. Contents Part I: Recombinant Protein Expression Into Escherichia Coli And Fermentation Conditions; Chapter 1: Introduction; Chapter 2: Construction Of Efficient Expression Vector (Plasmid); Chapter 3: Factors Affecting Transcription, Promoters, Upstream Elements, Transcriptional Terminators, Transcriptional Antitermin, Tightly Regulated Expression Systems; Chapter 4: Mrna Stability; Chapter 5: Factors Affecting Translation, Mrna Translational Initiator, Translational Enhancers, Translational Termination; Chapter 6: Expression Of Target Protein And The Compartments Of Expression, Cytoplasmic Expression, Periplasmic Expression, Extracellular Secretion; Chapter 7: Fusion Proteins; Chapter 8: Post-Translational Protein Folding; Chapter 8: Codon Usage; Chapter 10: Protein Degradation; Chapter 11: Fermentation Conditions For High-Density Cell Cultivation (Hdcc), Growth Medium, Efficient Production Of Recombinant Protein In Hdcc, Nutrient Feeding Strategy In Hdcc; Chapter 12: One Examples Of Protein Production Using E. Coli Expression System; Chapter 13: Conclusion. Part Ii: Recombinant Protein Expression Into Yeast, Pichia Pastoris And Fermentation Conditions; Chapter 1: Introduction; Chapter 2: Why P. Pastoris? Chapter 3: Construction Of Expression Strains, Expression Vectors, Alternative Promoters, Host Strains, Methanol Utilisation Phenotype, Protease-Reduced Host Strains, Integration Of Expression Vectors Into The P. Pastoris Genome, Generating Multicopy Strains; Chapter 4: Post-Translational Modifications Of Secreted Proteins, Secretion Signal Selection, N-Linked Glycosylation; Chapter 5: Production Of Recombinant Proteins In Fermenter Cultures Of The Yeast, Pichia Pastoris, Conceptual Basis For The P. Pastoris Expression System, High-Level Expression In Fermenter Cultures, Protein-Specific Adjustments To Improve Yield, Glycosylation Of Recombinant Proteins, Secretion Signals; Chapter 6: One Examples Of Protein Producing Using P. Pastoris Expression System, Chapter 7: Conclusion. Part Iii: Purification Strategies For Recombinant Proteins; Chapter 1: Purification Of Proteins; Chapter 2: Conventional Chromatography, Ion Exchange Chromatography, Reversed Phase Chromatography, Gel Permeation Chromatography, Affinity Chromatography, Affinity Tags, Cleavage, Conclusion. Part Iv: Market Size And Cost Analysis For The Production Of Therapeutic Proteins; Chapter 1: Market Size Of Therapeutic Proteins; Chapter 2: Outline Structure Of A Productin Unit And Cost Analysis For The Production Of Three Therapeutic Proteins. Part V: General Experimental Protocols; Chapter 1: Different Experimental Protocols, Preparation Of Genome Dna For E. Coli, A Differnt Method For Preparation Of Genomic Dna From Bacteria, Preparation Of Proteins From Periplasm (Osmotic Shock Method), Preparation Of Proteins From Outer Membrane, Transformation Of Plasmid Dna Into E. Coli (Calcium Chloride/Heat Shock Method), Transformation Of Plasmid Dna Into E. Coli (Electroporation), Sds-Page For Large Proteins, Sds-Page For Small Peptide, Pcr Amplification Of Dna, Protein Quantification: Brandford Method, Trans-Bloting For Protein, Restriction Enzyme Digestion Of Dna, Phenol/Chloroform Extraction Of Dna, Ethanol Precipitation Of Dna, Agarose Gel Electrophoresis, Transformation Of E. Coli By Electroporation (Alternative Method), Wizard Tm Pcr Preps Dna Purification System For Rapid, Purification Of Dna Fragments, Alternate Method For Purifying Dna From Agarose Gels, Southern Blotting, Rt Pcr Protocol, Using Superscript Reverse Transcriptase, Preparation Of Sequencing Gels, Isolation Of Rna From Mammalian Cells Using Rnazoltm (Teltest), Preparation For Yeast Transformation, Yeast Transformation, Digesting Prsq-Ura3 With Bamhi, Genomic Dna Preparation Of Yeast, Ligation (Circularisation) Of Genomic Dna Fragments, E. Coli Transformation (Alternate Method), Dna Miniprep From E. Coli (Alternate Method), Basic Plasmid Dna Isolation Protocol, Identification And Determination Of Amount Rec-Hum Proteins Via An Immunoenzymatic Test (Elisa), Determination Of Host Dna Contaminant Into R Hu Protein Through Dot Blot Method, Protocols For Down-Stream Processing.

Pichia Protocols

Pichia Protocols
Author: James M Cregg
Publisher: Springer Science & Business Media
Total Pages: 553
Release: 2007-08-08
Genre: Science
ISBN: 1588294293

This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.