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BioChemistry

Cloning Gene Expression and Protein Purification - Charles Hardin

Author

Charles Hardin
Jennifer Edwards
Andrew Riell
David Presutti, William Miller Dominique Robertson

Cover Price : Rs 995.00

Imprint : Oxford University Press
ISBN : 0195696530
YOP : 2008

Binding : Paperback
Total Pages : 448
CD : No

About the Book :- The central purpose of this combination lecture/laboratory manual is to present detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing the encoded protein, purifying the protein, and characterizing rudimentary aspects of its basic physical properties. The manual includes 20 experiments designed to train students to prepare, manipulate, and analyse plasmids, to produce fusion proteins in bacteria, and to purify these proteins based on unique chemical properties or substrate affinities. Electrophoresis, Southern and Western blotting and cominatorial techniques are emphasized. Radioisotopes, fluorophores and solvent effects on protein structure are discussed. (No radioisotopes are required.) The experiments included have been selected because of their high success rate and because they emphasize a project-oriented approach. Sufficient detail and background are provided such that the intended audience includes advanced undergraduate students, beginning graduates, and practising professionals engaged in a broad range of pursuits. Each experiment is accompanied by detailed process rationale. Where appropriate, alternate methods are also discussed. Maximizing its utility for both studetns and instructors, this unique manual precedes each lab experiment with the necessary background theory and principles, which culminate in a detailed process rationale and, ultimately, the experimental techniques themselves. While many lab manuals list background references, this innovative book combines both theory and procedure into one manageable volume. In addition to this theoretical background material, new innovations and insights are provided along with carefully selected primary source research literature. Contents :- Introductory Unit Introductory Lecture - Introduction to the Biochemical Laboratory Theory: Course Description Theory: "Central Dogma of Molecular Biology" Theory: Laboratory Safety Theory: The Scientific Method: Surviving Recipe Mentality Theory: Proactive Troubleshooting Theory: Introduction to the Biotechnology Laboratory Theory: Error Analysis and Assay Sensitivity Theory: Treatment of Analytical Data Theory: Concentration and Temperature Effects on pK a Introductory Lab 1 - Basic Biochemical Techniques I: Pipet Calibration and Solution Preparation Process: Pipets Introductory Lab 2 - Basic Techniques II: Absorbance Spectroscopy and Protein Concentration Determinations Process: AMP and Tryptophan Absorbance Spectra; Sample Calculations Theory: Absorption Data for the Nucleoside Monophosphates Process: Absorption Spectra Data for the Aromatic Amino Acids at pH 6; UV Absorption Characteristics of the Aromatic Amino Acids. Selected Extinction Coefficients Process: BCA Assay Sample Data Innov.: Measurement of Protein in 20 Seconds Part I - Nucleic Acids & Cloning Unit 1 Lecture 1 - DNA Isolation Theory: Subcloning Procedure Innov.: The pET Bacterial Plasmid System (Novagen) Lab 1.1 - Media Preparation; Bacterial Growths; Plasmid Minipreps; Hin dIII Digestion of DNA, Commercial Bacteriophage h DNA Bst EII Digest Size Standards Process: pUR278 and p2D Restriction Maps Process: Cloning the myo -3 Gene from C. elegans and Construction of an Expression Vector Process: C. elegans myo -3 Gene in pUR288 Vend. Lit.: Restriction Enzymes Hin dIII and Bst EII; h DNA Digests Process: Phage h Bst EII Digest Lab 1.2 - Agarose Gel Electrophoresis Exercises: Restriction Mapping Unit 2 Lecuture 2 - Construction of Recombinant Plasmids Innov.: Protecting and Manipulating Large DNA Substrates Innov.: Yeast of Burden--Yoking the YAC Lab 2.1 - Extraction and Cleanup of DNA Bands Cut from Agrose Gels, Quantitation of Yields and Ligation of myo -3 Hind III DNA Insert Fragment into Linearized B-gal Plasmid DNA Vend. Lit.: Gibco BRL (TM) T4 DNA Ligase Vend. Lit.: DNA Purification Kit (NaI/Glass Bead Method) Alt. App.: The Use of B-Agarase to Recover DNA from Gel Slices Alt. App.: GELase TM Unit 3 Lecture 3 - The Polymerase Chain Reaction Innov.: Polymerase Chain Reaction Used for Antigen Detection; Immuno-PCR: Very Sensitive Antigen Detection by Means of Specific Antibody-DNA Conjugates Lab 3.1 - Polymerase Chain Reaction Test for myo-3 Gene Insert Orientation Unit 4 Lecture 4 - Transcription of Genomic DNA & Analysis of the Resulting mRNAs Alt. App.: Isolation of Total RNA from E. coli Cells Alt. App.: Promega TM PolyATractTM System 1000 Alt. App.: Electrophoresis and Northern Blotting of RNA Unit 5 Lecture 5 - Transformation and Gene Expression Innov.: How Cells Respond to Stress Lab 5.1 - Preparation of Fresh Transformation - Competent Cells Alt. App.: Ultracomp TM Transformation Kit Lab 5.2 - Colony Immunoblotting to Screen for Transformants Alt. App.: The QIAexpressionist, QIAGENTM Unit 6 Lecture 6 - Analysis of DNA or RNA by Duplex Hybridization: DNA Isolation, Labeling, and Probing Innov.: Reduction of Background Problems in Nonradioactive Northern and Southern Blot Analyses Enables Higher Sensitivity than 32P-Based Hybridization Lab 6.1 - Labeling of DNA and Probe Construction from Cloned C. elegans myo-3 Gene; Quantitation of DNA Concentration Vend. Lit.: Digoxigenin Labeling of DNA: Genius TM Nucleic Acid Labeling System Lab 6.2 - Isolation of C. elegans Genomic DNA, Quantitation of DNA Concentration, and Digestion to Extract the myo-3 Gene Lab 6.3 - Southern Blotting Part 2 - Protein Purification Unit 7 Lecture 7 - Protein Purification Theory: Preparation and Handling of Biological Macromolecules for Crystallization Theory: Solution STructure of Biomacromolecules in Ionic Solutions Theory: Solubility as a Function of Protein Structure and Solvent Components Theory: Dominant Forces in Protein Folding Alt. App.: Hydrophobic Interaction Chromatography Alt. App.: Centriprep Microconcentrators for Small Volume Concentrations; Centricon-3 and 100 Lab 7.1 - The Protein Purifier: A Learning Aid from Pharmacia Lab 7.2 - Induction and Purification of B -Galactosidase Fusion Protein from Bacteria Lab 7.3 - Gel Filtration of Molecular Weight Standards and Protein Fractionation Process: Gel Filtration and Chromatography Vend. Lit.: Sephadex and Sephacryl Vend. Lit.: Sigma TM Gel Filtration Molecular Weight Markers Lab 7.4 - Mciroplate B -Galactosidase Assay to Determine Fractions Containing Fusion Protein; MW Determination Process: Time Course Assay of B-Galactosidase Vend. Lit.: B-Galactosidase Substrates Innov.: Luminescent Reporter Gene Assays for Luciferase and B-Galactosidase Using a Liquid Scintillation Counter Lab 7.5 - Ion Exchange Column Chromatography Process: Ion Exchange Chromatography Theory: The Isoelectric Point: Protein Charge Neutrality at a Particular pH Alt. App.: Ion-Pair Chromatography Alt. App.: HPLC: Ion Exchange and Reverse Phase Methods; Literature Sources Lab 7.6 - Affinity Chromatography and Microplate B -Galactosidase Assays to Determine Fractions Containing Fusion Protein Process: Affinity Chromatography Process: Affinity Chromatography: One Step Purification of Hybrid Proteins Carrying Fused B-Galactosidase Activity Lab 7.7 - BCA Protein Concentration Assays and B-Galactosidase Assays to Construct an Enzyme Purification Table Unit 8 Lecture 8 - Discontinuous Gel Electrophoresis, Protein Mobilities and Apparent Size Determination Process: Discontinuous Gel Electrophoresis & Protein Size Determination Lab 8.1 - Discontinuous SDS Gel Electrophoresis Unit 9 Lecture 9 - Immunochemical Techniques Innov.: Immunochemical Techniques Innov.: The Enzyme Linked Immunosorbent Assay (ELISA) Innov.: How the Immune System Learns About Self Innov.: Making Monoclonal Antibodies That Won't Fight Back Lab 9.1 - Western Blotting Process: Immunoblotting Process: Western Blots Using Stained Protein Gels Unit 10 Lecture 10 - Combinatorial Biochemical Technology Innov.: Examples of Combinatorial Techniques Innov.: Making Antibody Fragments Using Phage Display Libraries Innov.: Building a Better Enzyme Innov.: The ImmunoZAP Cloning and Expression System Appendices Part 1 Terms List Part 2 Terms List Laboratory Reagents Abbreviations List Copyright Acknowledgements Suggested Schedule Supplies Required Index Abbreviations: Innov., Innovation/Insight Theory, Theory/Principles Process, Process Rationale Vend. Lit., Vendor Literature Alt. App., Alternative Approach About the Authors :- Charles Hardin, Associate Professor of Biochemistry, North Carolina State University, Jennifer Edwards, Duke University, Andrew Riell, North Carolina State University, David Presutti, Adjunct Visiting Professor of Biochemistry, North Carolina State University, William Miller, William Neals Reynolds Professor of Biochemistry, North Carolina State University, and Dominique Robertson, Associate Professor of Botany, North Carolina State University.

Lecture Notes : Clinical Biochemistry - G J Beckett

Author

G J Beckett
Simon W Walker
Peter Rae
Peter Ashby

Cover Price : Rs 595.00

Imprint : Blackwell Science
ISBN : 1405154829
YOP : 2006
Edition : 2006

Binding : Paperback
Total Pages : 328
CD : No

DESCRIPTION Lecture Notes: Clinical Biochemistry presents the fundamental science underpinning common chemical investigations used in clinical practice. It takes a practically relevant approach that enables medical students and junior doctors to: Understand the value and limitations of chemical investigations Develop the skill to select appropriate investigations for diagnosis and management Interpret investigations correctly Develop a critical approach to diagnostic investigations Lecture Notes: Clinical Biochemistry takes a system-based approach. The underlying physiological rationale for any test is explained in the context of disruption by disease. This leads naturally to an integrated and practical understanding of chemical diagnostics. Case studies at the end of each chapter provide a clinical setting to help develop test-selection skills. Lecture Notes: Clinical Biochemistry provides the essential background to chemical investigations for medical students, junior doctors on foundation programmes, and nurses and practitioners involved in requesting or providing diagnostic service. CONTENTS Preface to the sixth edition List of abbreviations Chapter 1 Requesting and Interpreting Tests Chapter 2 Disturbances of Water Sodium and Potassium Balance Chapter 3 Acid-Base Balance and Oxygen Transport Chapter 4 Renal Disease Chapter 5 Disorders of Calcium, Magnesium and Phosphate Metabolism Chapter 6 Trauma, Inflammation, Immunity and Malignancy Chapter 7 Liver Disease Chapter 8 Gastrointestinal Tract Disease Chapter 9 Nutrition Chapter 10 Diabetes Mellitus and Hypoglycaemia Chapter 11 Cardiovascular Disorders Chapter 12 Disorders of Iron and Porphyrin Metabolism Chapter 13 Uric Acid, Gout and Purine Metabolism Chapter 14 Disorders of the Hypothalamus and Pituitary Chapter 15 Abnormailities of the Thyroid Function Chapter 16 Disorders of the Adrenal Cortex and Medulla Chapter 17 Investigation of Gonadal Function, Infertility, Menstrual Irregularities and Hirsutism Chapter 18 Pregnancy and Antenatal Screening Chapter 19 Clinical Biochemistry in Paediatrics and Geriatric Chapter 20 Central Nervous System and Cerebrospinal Fluid Chapter 21 Therapeutic Drug Monitoring and Chemical Toxicology Appendix Index. ABOUT THE AUTHORS Dr Geoff Beckett, Reader in Clinical Biochemistry at the University of Edinburgh Dr Simon Walker, Senior Lecturer in Clinical Biochemistry at the University of Edinburgh Dr Peter Rae, Consultant Clinical Biochemist, Western General Hospital, Edinburgh Dr Peter Ashby, Clinical Biochemist, Western General Hospital, Edinburgh

Blueprints Notes & Cases - Biochemistry, Genetics, and Embryology - Juan E Vargas

Author

Juan E Vargas
Annie Tan

Cover Price : Rs 425.00

Imprint : Blackwell Science
ISBN : 1405127252
YOP : 2004

Binding : Paperback
Total Pages : 192
CD : No

DESCRIPTION If you are wondering how the biochemistry principles you are studying will apply to real life patients, Blueprints Notes & Cases - Biochemistry, Genetics, and Embryology has just what you need – basic science concepts tied to clinical cases! This book offers high-yield, concise basic science content presented in a logical template. Each topic features a case presentation followed by thought questions and a basic science review. Thumbnails and key points provide a quick review of the essential information. Multiple-choice questions at the end of each case allow you to test your knowledge. Use during your coursework to aid in understanding application of principles, then review again as you prep for exams. Perfect for medical students -- physician assistants, nurse practitioners and related health professionals will also find Blueprints Notes & Cases valuable CONTENTS I. Biochemistry Carbohydrate and Lipid Metabolism 1. Glycolysis 2. TCA cycle 3. Gluconeogenesis 4. Glycogen Synthesis and Degradation 5. Oxidative Phosphorylation 6. Fatty Acid Metabolism Steroid and Cholesterol Metabolism 7. Steroid Biosynthesis 8. Cholesterol Synthesis 9. Cholesterol and Lipid Metabolism 10. Eicosanoid Biosynthesis Protein and Amino Acid Metabolism 11. Homocystinuria 12. Phenylketonuria 13. Collagen Synthesis 14. Purine Metabolism 15. Acute intermittent porphyria 16. Urea Cycle Cell Signaling and Coenzymes 17. IGF receptor signaling pathway 18. T-cell Receptor Pathways 19. G-proteins and Adenylate Cyclase 20. Vitamin Function II. Genetics Mendelian Genetics 21. Autosomal Dominant Inheritance 22. Autosomal Recessive Inheritance 23. X-Linked Recessive Inheritance 24. X-Linked Dominant Inheritance 25. Mitochondrial Inheritance 26. Variable Penetrance 27. Founder Effect Chromosomal Abnormalities 28. Chromosome Structure 29. Translocation 30. Sex Chromosomes 31. Gene Dosage Molecular Genetics 32. Genomic Imprinting 33. Viral Genetics 34. Gene Transcription and Translation 35. Immunoglobulin Genetics 36. Protein Structure and Function 37. DNA Repair 38. Cell Cycle III. Embryology 39. Embryonic Development 40. Fetal Development 41. Teratogens 42. Neural Development 43. Branchial Arch Anomalies 44. Cardiac Development 45. Pulmonary Development 46. Gastrointestinal Embryology 47. Renal Development

LIPID BIOCHEMISTRY - M I Gurr

Author

M I Gurr
J L Harwood
K N Frayn

Cover Price : Rs 595.00

Imprint : Blackwell Science
ISBN : 1405129190
YOP : 2004
Edition : 2004

Binding : Paperback
Total Pages : 320
CD : No

DESCRIPTION Since the publication of the first edition of this successful and popular book in 1970, the subject of lipid biochemistry has evolved greatly and this fifth up-to-date and comprehensive edition includes much new and exciting information. Lipid Biochemistry, fifth edition has been largely re-written in a user-friendly way, with chapters containing special interest topic boxes, summary points and lists of suggested reading, further enhancing the accessibility and readability of this excellent text. Contents include abbreviations and definitions used in the study of lipids, routine analytical methods, fatty acid structure and metabolism, dietary lipids and lipids as energy stores, lipid transport, lipids in cellular structures and the metabolism of structural lipids. The book provides a most comprehensive treatment of the subject, making it essential reading for all those working with or studying lipids. Upper level students of biochemistry, biology, clinical subjects, nutrition and food science will find the contents of this book invaluable as a study aid, as will postgraduates specializing in the topics covered in the book. Professionals working in research in academia and industry, including personnel involved in food and nutrition research, new product formulation, special diet formulation (including nutraceuticals and functional foods) and other clinical aspects will find a vast wealth of information within the book's pages. CONTENTS Preface Abbreviations Acknowledgement Lipids: definition, isolation, separation and detection Fatty acid structure and metabolism Lipids as energy stores Dietary lipids Lipid transport Lipids in cellular structures Metabolism of structural lipids Further reading Index . ABOUT THE AUTHORS Michael Gurr was a Visiting Professor in Human Nutrition at the University of Reading, UK and at Oxford Brookes University, UK. John Harwood is a Professor of Biochemistry at the School of Biosciences, Cardiff University, UK. Keith Frayn is a Professor of Human Metabolism at the Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, UK.

Plant Carbohydrate Biochemistry - J A Bryant

Author

J A Bryant
M M Burrell
N J Kruger

Cover Price : Rs 695.00

Imprint : T & F / Routledge
ISBN : 1859961126
YOP : 2004

Binding : Paperback
Total Pages : 316
CD : No

DESCRIPTION This book serves the urgent need to review the exciting developments and increasing research interest in this area, placing these within the context of our broader understanding of carbohydrate metabolism in higher plants. Plant Carbohydrate Biochemistry provides an integrated collection of definitive assessments, and synthesizes the disparate information appearing in primary research publications. The contributors to the book are established plant biochemists and physiologists chosen as much for their lucid exposition as for their scientific eminence. Plant Carbohydrate Biochemistry is dedicated to Professor T. ap Rees, a respected leader in the field of plant carbohydrate metabolism and respiration, as well as a dedicated and inspirational teacher. The book celebrates his achievements by focusing on recent advances in the areas of plant metabolism in which he had the greatest interest and influence as a researcher. It is therefore both a tribute to Professor ap Rees and a timely and authoritative statement of the current status of this subject. CONTENTS The first will be last and the last will be first: non-regulated enzymes call the tune? Modelling metabolic pathways and analysing control Carbohydrate: where does it come from, where does it go? The metabolism of sugars based upon sucrose Regulation of sucrose metabolism by protein phosphorylation: stimulation of sucrose synthesis by osmotic stress and 5-aminoimidazole-4-carboxamide riboside Sugars, far more that just fuel for plant growth The synthesis of the starch granule From enzyme activity to flux control: a quest to understand starch deposition in developing cereal grains Allosteric regulation and reductive activation of ADPglucose pyrophosphorylase Regulation of rubisco Biochemistry of photorespiration and the consequences for plant performance The use of mutants of amaranthus edulis to study carbon and nitrogen metabolism in C4 photosynthesis On being thick: fathoming apparently futile pathways of photosynthesis and carbohydrate metabolism in succulent CAM plants Phosphoenolpyruvate carboxykinase in plants: its role and regulation Ascorbic acid metabolism in plants Implications of inter - and intracellular compartmentation for the movement of metabolites in plant cells Metabolite transporters in plastids and their role in photoassimilate allocation Starch synthesis in plant storage tissues: precursor dependency and function of the plastidic ATP/ADP transporter The folate status of plant mitochondria Compartmentation of tetrapyrrole synthesis in plant cells What's a nice enzyme like you doing in a place like this? A possible link between glycolysis and DNA replication.

Bio-Nano-Geo Sciences : The Future Challenge - Alok Srivastava

Author

Alok Srivastava
Ipsita Roy

Cover Price : Rs 795.00

Imprint : Ane Books Pvt. Ltd.
ISBN : 818052180x
YOP : 2009

Binding : Hardback
Total Pages : 256
CD : No

About the Book :- The book covers topics as varied as Natural Nano-Machines of Life: A Biological Route to Nanotechnology, River Response to Climate Change, Ethics in Public Domain: Biomedical Research and Beyond, Arsenicosis and the Arsenic Selenium Connection, Pharmacoscintigraphic evaluation of Nanoparticle Drug Delivery Systems besides articles covering a number of other interesting facets of the above mentioned subject area. Contents :- 1. Natural Nano-Machines of Life: A Biological Route to Nanotechnology 2. Hybrid Bio-Nanodevices for Engineering and Healthcare 3. Pharmacoscintigraphic Evaluation of Nanoparticulate Drug Delivery Systems 4. Nano Materials of Carbon – Fullerenes and Nanotubes 5. Synthesis and Characterization of Nanosized Materials for Lithium Rechargeable Batteries 6. Clay Nano-composites as Environmental Friendly Catalysts for Green Chemical Synthesis 7. Micronutrient Disorders – Challenge to Sustain Food Grain Production 8. Using Plants for Remediation of Metal Polluted Soils 9. Pollution in the Red- Bloomed Himalayan Dal Lake of Kashmir 10. Production of Radioisotopes for Positron Emission Tomography 11. Study of Radiotracer Applications in Bio-Nano-Geo Sciences at Saha Institute of Nuclear Physics 12. Arsenicosis and the Arsenic-Selenium Connection 13. Biogeochemistry of Lacustrine Sediment: An Archive of Paleoclimate Variablity 14. Engineering Interventions for Management of Marginally Saline Groundwaters 15. River Response to Climate Change: Why, Where and How? 16. Recession Rate of Tributary of Gangotri Glacier System, Garhwal Himalaya-India 17. Molecular Technologies Application in Cancer Diagnosis for Improved Treatment Strategy 18. Metagenomics: A Splendid Novel Approach to Explore and Exploit Microbial Diversity 19. Ethics in Public Domain: Biomedical Research and Beyond. About the Editors :- Dr. Alok Srivastava, a Nuclear Chemist, is presently teaching in the Chemistry Department of Panjab University, Chandigarh. He did his doctorate in 1985 from the Radiochemistry Division of Bhabha Atomic Research Centre, Mumbai in the field of Low Energy Nuclear Fission. He was a Humboldt Fellow in the Institute of Nuclear Chemistry, University of Mainz, Germany in 1986-88. Dr.Srivastava was invited to work as a DAAD Professor in the Aachen University of Applied Sciences, Germany for two years in the period 2002-04. He has been a Consultant to EURATOM which is one of the Scientific Bodies of the European Community. He presently works in the area of Material Science, Environmental Science and Nuclear Reactions. Dr. Ipsita Roy received her Ph.D in Applied Biochemistry from the IIT, Delhi. She worked there as a Project Scientist before moving on to the University of Bonn, Germany as an Alexander von Humboldt Fellow. She joined the Department of Biotechnology, National Institute of Pharmaceutical Education and Research in Mohali in 2005 where she works as an Assistant Professor. She is involved in the teaching of post graduate pharmacy students as well as conducting research in Biotechnology. Her areas of interest include Protein stabilization using small molecules and aptamers.

 

Author

Michael M. Cox
David L. Nelson

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