Split Inteins PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Split Inteins PDF full book. Access full book title Split Inteins by Henning D. Mootz. Download full books in PDF and EPUB format.
Author: Henning D. Mootz
Publisher:
ISBN: 9781493964512
Category : Amino acid sequence
Languages : en
Pages : 270
Get Book Here
Book Description
"This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments."--OCLC.
Author: Henning D. Mootz
Publisher:
ISBN: 9781493964512
Category : Amino acid sequence
Languages : en
Pages : 270
Get Book Here
Book Description
"This volume focuses on applications of split inteins, and the progress that has been made in the past 5 years on discovery and engineering of fast and more efficient split inteins. The first few chapters in Split Inteins: Methods and Protocols explore new techniques on how to use split inteins for affinity purification of overproduced proteins, and split-intein based technologies to prepare cyclic peptides and proteins. The next few chapters discuss semisynthetic protein trans-splicing using one synthetic intein piece, synthetic intein-extein pieces used to deliver other cargos for chemical modification both of purified proteins and of proteins in living cells, as well as isotopic labeling of proteins for NMR studies, and a discussion on how protein block copolymers can be generated by protein trans-splicing to form protein hydrogels. The last few chapters deal with intein applications in transgenic plants and conditional inteins that can be regulated in artificial ways by small molecules or light, a cassette-based approach to quickly test many intein insertion positions, and a computational approach to predict new intein split sites (the approach also works for other proteins). Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Split Inteins: Methods and Protocols is a valuable resource that will provide guidance toward possibilities of split intein applications, explore proven and detailed protocols adaptable to various research projects, and inspire new method developments."--OCLC.
Author: Miquel Vila-Perelló
Publisher: Humana
ISBN: 9781071604335
Category : Science
Languages : en
Pages : 0
Get Book Here
Book Description
This book provides a comprehensive overview of Expressed Protein Ligation (EPL), detailing methods and protocols to generate site-specifically modified proteins. Chapters include an overview of the protein semi-synthesis field, as well as related areas that have contributed to the development of EPL such as protein splicing and peptide synthesis. Following the introductory chapters, the rest of the book guides readers through protocols to perform EPL reactions, methods to synthesize peptide thioesters and to perform peptide and protein ligations, label proteins inside living cells, protocols for the semi-synthesis of phorphorylated, glycosylated and ubiquitylated proteins, synthesis and assembly of assymetrically modified nucleosomes, use of ligation auxiliaries and synthesis of cyclic proteins, as well as novel desulfurization strategies and use of selective Cys side chain protection to obtain precisely modified proteins.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 tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Expressed Protein Ligation: Methods and Protocols will ensure successful implementation of protein semi-synthesis methods to further study the structure and function of proteins.
Author: Marlene Belfort
Publisher: Springer Science & Business Media
ISBN: 9783540852353
Category : Science
Languages : en
Pages : 404
Get Book Here
Book Description
This book provides the first and only comprehensive description and detailed summary of the genetics, structure, function, mechanisms of action, evolution and engineering of homing endonucleases and inteins. These two unique protein superfamilies, which are tied together through their frequent fusion and coevolution, have generated considerable excitement for their fundamental, structural, and functional properties, their evolution as parasitic elements, and their widespread applications as gene targeting agents and as instruments for the generation of modified proteins and novel protein combinations.
Author: Luca D. D'Andrea
Publisher: John Wiley & Sons
ISBN: 1119044103
Category : Science
Languages : en
Pages : 492
Get Book Here
Book Description
Presenting a wide array of information on chemical ligation – one of the more powerful tools for protein and peptide synthesis – this book helps readers understand key methodologies and applications that protein therapeutic synthesis, drug discovery, and molecular imaging. • Moves from fundamental to applied aspects, so that novice readers can follow the entire book and apply these reactions in the lab • Presents a wide array of information on chemical ligation reactions, otherwise scattered across the literature, into one source • Features comprehensive and multidisciplinary coverage that goes from basics to advanced topics • Helps researchers choose the right chemical ligation technique for their needs
Author: Robert E. Rhoads
Publisher: Humana
ISBN: 9781493936236
Category : Medical
Languages : en
Pages : 0
Get Book Here
Book Description
This volume presents detailed laboratory protocols for in vitro synthesis of mRNA with favorable properties, its introduction into cells by a variety of techniques, and the measurement of physiological and clinical consequences such as protein replacement and cancer immunotherapy. Synthetic techniques are described for structural features in mRNA that provide investigational tools such as fluorescence emission, click chemistry, photo-chemical crosslinking, and that produce mRNA with increased stability in the cell, increased translational efficiency, and reduced activation of the innate immune response. Protocols are described for clinical applications such as large-scale transfection of dendritic cells, production of GMP-grade mRNA, redirecting T cell specificity, and use of molecular adjuvants for RNA vaccines. 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Synthetic mRNA: Production, Introduction into Cells, and Physiological Consequences is a valuable and cutting-edge resource for both laboratory investigators and clinicians interested in this powerful and rapidly evolving technology.
Author: Yamin Fan (Ph. D. in chemical engineering)
Publisher:
ISBN:
Category : Bioconjugates
Languages : en
Pages :
Get Book Here
Book Description
Over the past two decades, inteins have been extensively used in a wide variety of applications in biotechnology. Split inteins are a subset of inteins, which are identified more recently and expressed in two separate segments naturally. They catalyze the splicing reaction in trans upon association of the two halves. Due to their unique features, split inteins offer improved controllability and flexibility in trans-splicing and trans-cleaving over the previous tools based on contiguous inteins. The engineered split inteins would allow the development of efficient self-cleaving affinity tags for purification applications and new methods for protein conjugation. In this work, an engineered split intein derived from Nostoc punciforme (Npu) was applied in a column-free purification strategy in combination with the aggregating tag, elastin-like polypeptide (ELP), as an initial capture step for recombinant proteins expressed in E. coli. Meanwhile, on-column purification strategy using the same engineered split intein was employed for the production of value-added biosimilar target, Granulocyte-colony stimulating factor (G-CSF). To adapt the split intein-based purification platform for the production of protein therapeutics expressed in mammalian cells, multiple leader sequences were designed and screened for optimal expression and secretion of intein-tagged precursor proteins. Moreover, the extein dependency of this engineered Npu split intein was thoroughly characterized by using in solution cleaving kinetics study and Forster Resonance Energy Transfer (FRET) based high-throughput method. The information gathered guides for fast and consistent cleavage reactions among various target proteins and provides insight to the cleavage mechanism. In this work, the trans-splicing properties of split inteins were also exploited for developing novel bioconjugation methods onto fluorescent nanodiamonds. Two split inteins, Gos-TerL and GP41.1, were used for the development of N-terminal and C-terminal oriented bioconjugation schemes, respectively. The new methods would allow rapid and spontaneous immobilization of proteins onto fluorescent nanodiamond surfaces for applications such as biomedical imaging or drug delivery.
Author: Arnaud Gautier
Publisher: Humana Press
ISBN: 9781493922710
Category : Science
Languages : en
Pages : 0
Get Book Here
Book Description
This detailed volume provides in-depth protocols for protein labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. Chapters provide protocols for labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Site-Specific Protein Labeling: Methods and Protocols provides a comprehensive overview on the most relevant and established labeling methodologies, and helps researchers to choose the most appropriate labeling method for their biological question.
Author: Richard G. Weiss
Publisher: Springer Science & Business Media
ISBN: 1402036892
Category : Science
Languages : en
Pages : 969
Get Book Here
Book Description
"Molecular Gels: Materials with Self-Assembled Fibrillar Networks" is a comprehensive treatise on gelators, especially low molecular-mass gelators and the properties of their gels. The structures and modes of formation of the self-assembled fibrillar networks (SAFINs) that immobilize the liquid components of the gels are discussed experimentally and theoretically. The spectroscopic, rheological, and structural features of the different classes of low molecular-mass gelators are also presented. Many examples of the application of the principal analytical techniques for investigation of molecular gels (including SANS, SAXS, WAXS, UV-vis absorption, fluorescence and CD spectroscopies, scanning electron, transmission electron and optical microscopies, and molecular modeling) are presented didactically and in-depth, as are several of the theories of the stages of aggregation of individual low molecular-mass gelator molecules leading to SAFINs. Several actual and potential applications of molecular gels in disparate fields (from silicate replication of nanostructures to art conservation) are described. Special emphasis is placed on perspectives for future developments. This book is an invaluable resource for researchers and practitioners either already researching self-assembly and soft matter or new to the area. Those who will find the book useful include chemists, engineers, spectroscopists, physicists, biologists, theoreticians, and materials scientists.
Author: Stanley Wong
Publisher:
ISBN:
Category :
Languages : en
Pages :
Get Book Here
Book Description
Author:
Publisher: Academic Press
ISBN: 0128191295
Category : Science
Languages : en
Pages : 310
Get Book Here
Book Description
Chemical and Synthetic Biology Approaches to Understand Cellular Functions - Part C, Volume 633, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial. This release includes sections on Next generation probes for molecular imaging in cells, Competitive binding assay for biotin and biotin derivatives, based on avidin and biotin-4-fluorescein, Converting avidin to bind ligands other than biotin, especially steroids, Chemoenzymatic Labeling Strategy, Engineered Siderophores, Small molecules to inhibit bacterial population behavior, NMR tube bioreactor, Small molecule controlled RAS activation system, Small molecule regulated Cas9, the Design and application of synthetic receptors, and much more. - Contains the authority of authors who are leaders in their field - Provides a comprehensive source on new methods and research in enzymology
