|
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
|
|
Microsoft VBScript runtime error '800a000d'
Type mismatch: 'formatnumber'
/detailed-booklist.asp, line 239 | | |