ProxiMAX Randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

ProxiMAX randomisation achieves saturation mutagenesis of contiguous codons without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, it uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents and as such, requires no specialised chemistry, reagents nor equipment. When particular residues are known to affect protein activity/specificity, their combinatorial replacement with all 20 amino acids, or a subset thereof, can provide a rapid route to generating proteins with desirable characteristics. Conventionally, saturation mutagenesis replaced key codons with degenerate ones. Although simple to perform, that procedure resulted in unnecessarily large libraries, termination codons and inherent uneven amino acid representation. ProxiMAX randomisation is an enzyme-based technique that can encode unbiased representation of all or selected amino acids or else can provide required codons in pre-defined ratios. Each saturated position can be defined independently of the others. ProxiMAX randomisation is achieved via saturation cycling: an iterative process comprising blunt end ligation, amplification and digestion with a Type IIS restriction enzyme. We demonstrate both unbiased saturation of a short 6-mer peptide and saturation of a hypervariable region of a scfv antibody fragment, where 11 contiguous codons are saturated with selected codons, in pre-defined ratios. As such, ProxiMAX randomisation is particularly relevant to antibody engineering. The development of ProxiMAX randomisation from concept to reality is described.

Casebook on contract law

Jill Poole's bestselling Casebook on Contract Law provides students with a comprehensive selection of case law which addresses all aspects of the subject encountered on undergraduate courses. Extracts have been chosen from a wide range of historical and contemporary cases to illustrate the reasoning processes of the courts and to show how legal principles are developed. The cases can be analysed and discussed independently while, taken as a whole, the chapters provide a sound understanding of the modern law of contract. Key features: * An introductory chapter provides valuable guidance on how to use a casebook to help you develop the essential skill of reading and analysing cases * Cases are accompanied by succinct author commentary designed to highlight the key elements of each case * The two colour text design distinguishes case extracts from author commentary ensuring the book is easy to use and navigate through The casebook is fully supported by an Online Resource Centre, which provides: * Exercises and guidance on reading cases * Self-test questions and answers * Guidance on how to answer problem-style questions * Updates

Casebook on contract law

Jill Poole's best-selling Casebook on Contract Law provides a clear and well-structured explanation of the principles and rules of contract law through a comprehensive selection of case law, addressing all aspects encountered on undergraduate courses. The coverage in this new edition has been revised to incorporate all recent significant decisions and judgments made by the House of Lords and the Court of Appeal. Extracts have been chosen from a wide range of historical and contemporary cases to illustrate the reasoning processes of the court and how legal principles are developed, thus enabling cases to be analysed and discussed independently while, taken as a whole, the chapters provide a sound understanding of the modern law of contract. Succinct author commentary focuses the reader on the key elements within the extracts, while thought-provoking questions are posed throughout to develop a more in-depth appreciation of the subject. Online resource centre Student resources - Updates - Guidance on answering problem-style questions - Exercises and guidance on reading cases - Self test questions and answers - 'Ask the author' section

Textbook on contract law

Jill Poole's immensely popular Textbook on Contract Law has been guiding students through contract law for many years. The accessible writing style and focus on key principles and developments in contract law make this text a favourite with students and lecturers alike. The author places the law of contract clearly within its wider context before proceeding to provide detailed yet accessible treatment of all the key areas encountered when studying contract law. Key features: · Key points, illustrative examples and questions encourage a deeper understanding of the central facts and issues · Each chapter begins with a summary of key issues, providing an overview of key themes and points of law, and concludes with suggestions for further reading, guiding you towards the most relevant texts and articles · An attractive page design highlights headings, case summaries, case extracts and a variety of learning features, making navigation through the text easy Fully supported by an Online Resource Centre, which provides: Student Resources Regular updates on new legislation, cases, and other important developments Guidance on answering questions in contract law Self-test questions and answers linked with Casebook on Contract Law Lecturer Resources A test bank of 300 multiple choice questions with answers and feedback

Textbook on contract law

This tenth edition of the established Textbook on Contract Law by Jill Poole provides a wide-ranging and straightforward exposition of contract law. The text opens with an overview of the main issues surrounding contract law, which places the subject in its wider context, then goes on to give a clear explanation of all the major areas of contract law encountered on undergraduate courses. Features of the book include chapter summaries to draw key themes and issues together; examples and questions to encourage a deeper understanding of the often complex points of law; and extensive further reading lists of both texts and articles to guide students towards the most relevant and up-to-date resources available. Online resource centre Lecturer resources - Testbank of multiple choice questions Student resources - Guidance on answering problem-style questions in contract law - Self test questions and answers - Student questions - Updates - 'Ask the Author' section

Contract law concentrate:law revision and study guide

Contract Law Concentrate is a high quality revision guide which covers the main topics found on undergraduate courses. The clear, succinct coverage of key legal points within a specific topic area, including key cases, enables students to quickly grasp the fundamental principles of Contract law. Written by Jill Poole, an experienced teacher and examiner and author of Textbook on Contract Law and Casebook on Contract law. The book focuses on the needs of students to pass their exams with a number of pedagogical features which help with the preparation for exams and suggest ways to improve marks. Endorsed by students and lecturers for level of coverage, accuracy and exam advice. Online Resource Centre Interactive flashcards Glossary Exam and revision guidance.

Textbook on contract law

This edition of the guide to the law of contract takes account of the implications of Internet contracting and includes discussion of the Consumer Protection (Distance Selling) Regulations 2000. Where appropriate, reference is made, for reasons of comparison, to the principles contained in thePrinciples of European Contract Law (PECL) and the UNIDROIT Principles of International Commercial Contracts.;Significant developments in contract law, both statutory and case law, are discussed, including the Contracts (Rights of Third Parties) Act 1999, the Electronic Communications Act 2000 andthe House of Lords' decisions in Alfred McAlpine Construction Ltd v Panatown Ltd and Attorney General V Blake. Other important decisions covered in this revised edition are Royal Bank of Scotland v Etridge, Barclays Bank Plc v Coleman, Barclays Bank Plc v Boulter, Avon Insurance v Swire, Zanzibar vBritish Aerospace (Lancsaster House) Ltd and Nutt v Read. In addition, there is discussion of the Unfair Terms in Consumer Contracts Regulations 1999 and the European Commission's Review of the Unfair Terms Directive, as well as coverage of Director General of Fair Trading v First National Bank. Other decisions on illegality, onerous terms, constructionand repudiation of contract are included.

Textbook on contract law

Now in its 12th edition, Jill Poole's immensely popular Textbook on Contract Law has been guiding students through contract law for many years. The accessible writing style and focus on key principles and developments in contract law make this text a favourite with students and lecturers alike. The author places the law of contract clearly within its wider context before proceeding to provide a detailed yet accessible treatment of all the key areas encountered when studying contract law. Key features: -Each chapter begins with a summary of key issues, providing an overview of key themes and points of law, and concludes with suggestions for further reading, guiding you towards the most relevant texts and articles -Key points, illustrative examples and questions encourage a deeper understanding of the central facts and issues -Headings, case summaries and case extract boxes allow for easy navigation through the text

Casebook on contract law

•The introductory chapter provides valuable guidance on how to use a casebook to help develop the essential skills of reading and analysing cases • Cases are accompanied by succinct author commentary designed to highlight the key elements of each case • Thought-provoking questions are posed throughout to develop an in-depth understanding of the subject •The book is easy to use and navigate through as the case extracts are clearly distinguished from author commentary Jill Poole's bestselling Casebook on Contract Law provides students with a comprehensive selection of case law which addresses all aspects of the subject encountered on undergraduate courses. Extracts have been chosen from a wide range of historical and contemporary cases to illustrate the reasoning processes of the courts and to show how legal principles are developed. The cases can either be analysed and discussed independently or, taken as a whole, the cases form chapters that provide a sound understanding of the modern law of contract. The casebook is fully supported by an Online Resource Centre, which provides: -Exercises and guidance on reading cases -Guidance on answering questions in contract law -Self-test questions and answers

ProxiMAX randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

Back in 2003, we published ‘MAX’ randomisation, a process of non-degenerate saturation mutagenesis using exactly 20 codons (one for each amino acid) or else any required subset of those 20 codons. ‘MAX’ randomisation saturates codons located in isolated positions within a protein, as might be required in enzyme engineering, or else on one face of an alpha-helix, as in zinc finger engineering. Since that time, we have been asked for an equivalent process that can saturate multiple, contiguous codons in a non-degenerate manner. We have now developed ‘ProxiMAX’ randomisation, which does just that: generating DNA cassettes for saturation mutagenesis without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, ProxiMAX randomisation uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents. Thus it requires no specialised chemistry, reagents nor equipment and simply relies on a process of saturation cycling comprising ligation, amplification and digestion for each cycle. The process can encode both unbiased representation of selected amino acids or else encode them in pre-defined ratios. Each saturated position can be defined independently of the others. We demonstrate accurate saturation of up to 11 contiguous codons. As such, ProxiMAX randomisation is particularly relevant to antibody engineering.

NOx storage-reduction characteristics of Ba-based lean NOx trap catalysts subjected to simulated road aging

In order to study the effect of washcoat composition on lean NOx trap (LNT) aging characteristics, fully formulated monolithic LNT catalysts containing varying amounts of La-stabilized CeO2 (5 wt% La2O3) or CeO2-ZrO2 (Ce:Zr = 70:30) were subjected to accelerated aging on a bench reactor. Subsequent catalyst evaluation revealed that aging resulted in deterioration of the NOx storage, NOx release and NOx reduction functions, whereas the observation of lean phase NO2 slip for all of the aged catalysts indicated that LNT performance was not limited by the kinetics of NO oxidation. After aging, all of the catalysts showed increased selectivity to NH3 in the temperature range 250–450 °C. TEM, H2 chemisorption, XPS and elemental analysis data revealed two main changes which can explain the degradation in LNT performance. First, residual sulfur in the catalysts, present as BaSO4, decreased catalyst NOx storage capacity. Second, sintering of the precious metals in the washcoat was observed, which can be expected to decrease the rate of NOx reduction. Additionally, sintering is hypothesized to result in segregation of the precious metal and Ba phases, resulting in less efficient NOx spillover from Pt to Ba during NOx adsorption, as well as decreased rates of reductant spillover from Pt to Ba and reverse NOx spillover during catalyst regeneration. Spectacular improvement in LNT durability was observed for catalysts containing CeO2 or CeO2-ZrO2 relative to their non-ceria containing analog. This was attributed to (i) the ability of ceria to participate in NOx storage/reduction as a supplement to the main Ba NOx storage component; (ii) the fact that Pt and CeO2(-ZrO2) are not subject to phase segregation; and (iii) the ability of ceria to trap sulfur, resulting in decreased sulfur accumulation on the Ba component.

Contract law concentrate

Contract Law Concentrate is a high quality revision guide which covers the main topics found on undergraduate courses. The clear, succinct coverage of key legal points within a specific topic area, including key cases, enables students to quickly grasp the fundamental principles of contract law. Written by Jill Poole, an experienced teacher and examiner and author of Textbook on Contract Law and Casebook on Contract Law, the book focuses on the needs of students to pass their exams. A number of pedagogical features help with the preparation for exams and suggest ways to improve marks. This guide has been rigorously reviewed and is endorsed by students and lecturers for level of coverage, accuracy, and exam advice. Packed with essential information, key cases, revision tips, exam Q&As, and more, Contract Law Concentrate is also supported by extensive online resources to take your learning further (www.oxford.com/lawrevision/): · Test your knowledge with the multiple choice questions and receive feedback on your answers. · Revise the facts and discussions of key cases using the interactive flashcards. · Learn the important terms and definitions using the interactive glossary. · Check that you have covered the main points of a topic using the key facts lists. · Achieve better marks following the advice on revision and exam technique by experienced examiner Nigel Foster.

The integration of ProxiMAX randomisation with CIS display for the production of novel peptides

Saturation mutagenesis is a powerful tool in modern protein engineering, which permits key residues within a protein to be targeted in order to potentially enhance specific functionalities. However, the creation of large libraries using conventional saturation mutagenesis with degenerate codons (NNN or NNK/S) has inherent redundancy and consequent disparities in codon representation. Therefore, both chemical (trinucleotide phosphoramidites) and biological methods (sequential, enzymatic single codon additions) of non-degenerate saturation mutagenesis have been developed in order to combat these issues and so improve library quality. Large libraries with multiple saturated positions can be limited by the method used to screen them. Although the traditional screening method of choice, cell-dependent methods, such as phage display, are limited by the need for transformation. A number of cell-free screening methods, such as CIS display, which link the screened phenotype with the encoded genotype, have the capability of screening libraries with up to 1014 members. This thesis describes the further development of ProxiMAX technology to reduce library codon bias and its integration with CIS display to screen the resulting library. Synthetic MAX oligonucleotides are ligated to an acceptor base sequence, amplified, and digested, subsequently adding a randomised codon to the acceptor, which forms an iterative cycle using the digested product of the previous cycle as the base sequence for the next. Initial use of ProxiMAX highlighted areas of the process where changes could be implemented in order to improve the codon representation in the final library. The refined process was used to construct a monomeric anti-NGF peptide library, based on two proprietary dimeric peptides (Isogenica) that bind NGF. The resulting library showed greatly improved codon representation that equated to a theoretical diversity of ~69%. The library was subsequently screened using CIS display and the discovered peptides assessed for NGF-TrkA inhibition by ELISA. Despite binding to TrkA, these peptides showed lower levels of inhibition of the NGF-TrkA interaction than the parental dimeric peptides, highlighting the importance of dimerization for inhibition of NGF-TrkA binding.

Stomach content records for pelagic fish (herring, mackerel, blue whiting, albacore and bluefin tuna) in the northeast Atlantic

Fish stomach content records extracted from the DAPSTOM 4.5 database (held at the UK Centre for Environment, Fisheries and Aquaculture Science - CEFAS). Data collated as part of the EU Euro-Basin project and specifically concerning herring (Clupea harengus), mackerel (Scomber scombrus), blue whiting (Micromesistius poutassou), albacore (Thunnus alalunga) and bluefin tuna (Thunnus thynnus). The data set consist of 20720 records - collected throughout the northeast Atlantic, between 1906 and 2011 - mostly during routine fisheries monitoring research cruises.