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Author: Robert Damoiseaux
Publisher: Humana Press
ISBN: 9781493977222
Category : Medical
Languages : en
Pages : 234
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Book Description
This volume explores the applications of reporter gene technology and the methodologies needed for their effective implementation. The chapters in this book cover practical topics such as how to integrate reporter constructs into cellular models, viral delivery, splicing applications, in vivo imaging, and a guide to the use of multi-cistronic constructs. Additionally, chapters also include detailed mechanistic uses of reporter genes in cellular pathways, and a look at project and data management of screening applications. 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. Cutting-edge and thorough, Reporter Gene Assays: Methods and Protocols is a valuable resource for anyone who is interested in learning more about reporter genes.
Author: Robert Damoiseaux
Publisher: Humana Press
ISBN: 9781493977222
Category : Medical
Languages : en
Pages : 234
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Book Description
This volume explores the applications of reporter gene technology and the methodologies needed for their effective implementation. The chapters in this book cover practical topics such as how to integrate reporter constructs into cellular models, viral delivery, splicing applications, in vivo imaging, and a guide to the use of multi-cistronic constructs. Additionally, chapters also include detailed mechanistic uses of reporter genes in cellular pathways, and a look at project and data management of screening applications. 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. Cutting-edge and thorough, Reporter Gene Assays: Methods and Protocols is a valuable resource for anyone who is interested in learning more about reporter genes.
Author: Manfred Schwab
Publisher: Springer Science & Business Media
ISBN: 3540368477
Category : Medical
Languages : en
Pages : 3307
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Book Description
This comprehensive encyclopedic reference provides rapid access to focused information on topics of cancer research for clinicians, research scientists and advanced students. Given the overwhelming success of the first edition, which appeared in 2001, and fast development in the different fields of cancer research, it has been decided to publish a second fully revised and expanded edition. With an A-Z format of over 7,000 entries, more than 1,000 contributing authors provide a complete reference to cancer. The merging of different basic and clinical scientific disciplines towards the common goal of fighting cancer makes such a comprehensive reference source all the more timely.
Author: Mousumi Debnath
Publisher: Springer Science & Business Media
ISBN: 9048132614
Category : Medical
Languages : en
Pages : 527
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Book Description
A rapid development in diverse areas of molecular biology and genetic engineering resulted in emergence of variety of tools. These tools are not only applicable to basic researches being carried out world over, but also exploited for precise detection of abnormal conditions in plants, animals and human body. Although a basic researcher is well versed with few techniques used by him/her in the laboratory, they may not be well acquainted with methodologies, which can be used to work out some of their own research problems. The picture is more blurred when the molecular diagnostic tools are to be used by physicians, scientists and technicians working in diagnostic laboratories in hospitals, industry and academic institutions. Since many of them are not trained in basics of these methods, they come across several gray areas in understanding of these tools. The accurate application of molecular diagnostic tools demands in depth understanding of the methodology for precise detection of the abnormal condition of living body. To meet the requirements of a good book on molecular diagnostics of students, physicians, scientists working in agricultural, veterinary, medical and pharmaceutical sciences, it needs to expose the reader lucidly to: Give basic science behind commonly used tools in diagnostics Expose the readers to detailed applications of these tools and Make them aware the availability of such diagnostic tools The book will attract additional audience of pathologists, medical microbiologists, pharmaceutical sciences, agricultural scientists and veterinary doctors if the following topics are incorporated at appropriate places in Unit II or separately as a part of Unit-III in the book. Molecular diagnosis of diseases in agricultural crops Molecular diagnosis of veterinary diseases. Molecular epidemiology, which helps to differentiate various epidemic strains and sources of disease outbreaks. Even in different units of the same hospital, the infections could be by different strains of the same species and the information becomes valuable for infection control strategies. Drug resistance is a growing problem for bacterial, fungal and parasitic microbes and the molecular biology tools can help to detect the drug resistance genes without the cultivation and in vitro sensitivity testing. Molecular diagnostics offers faster help in the selection of the proper antibiotic for the treatment of tuberculosis, which is a major problem of the in the developing world. The conventional culture and drug sensitivity testing of tuberculosis bacilli is laborious and time consuming, whereas molecular diagnosis offers rapid drug resistant gene detection even from direct clinical samples. The same approach for HIV, malaria and many more diseases needs to be considered. Molecular diagnostics in the detection of diseases during foetal life is an upcoming area in the foetal medicine in case of genetic abnormalities and infectious like TORCH complex etc. The book will be equally useful to students, scientists and professionals working in the field of molecular diagnostics.
Author: Sean R. Gallagher
Publisher: Academic Press
ISBN: 0323137644
Category : Science
Languages : en
Pages : 250
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Book Description
The gb-glucuronidase (GUS) gene is extremely useful as a reporter of the expression of introduced genes and can be used in organisms where other reporter genes are useless. Thus, the GUS gene is the reporter gene of choice for transgenic plant research. Not only can this assay be used to detect whether a gene is being expressed, but it can be used to determine the location of the gene product within cells. Low cost is another advantage of this assay. GUS Protocols provides instructions and essential background information that will enable researchers to effectively use the GUS gene as a reporter of the expression of introduced genes. - First book on the GUS reporter system - Up-to-date reference lists following each of the fourteen chapters - Comb-bound for convenient bench-top use - Written by leading authorities including R.A. Jefferson, inventor of the GUS assay - Illustrated with color - GUS detection by fluorometric, spectrophotometric, and histochemical methods - Fast, automated assays
Author: OECD
Publisher: OECD Publishing
ISBN: 9264122818
Category :
Languages : en
Pages : 16
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Book Description
This Test Guideline describes an in vivo assay that detects chemicals that may induce gene mutations. In this assay, transgenic rats or mice that contain multiple copies of chromosomally integrated plasmid or phage shuttle vectors are used. The ...
Author: You Li
Publisher:
ISBN:
Category : Biological assay
Languages : en
Pages : 0
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Book Description
Traditional cell-based assays have a high failure rate of identifying effective compounds in drug discovery. To address this problem, recent drug research has focused on engineering cancer cells that can indicate specific drug molecular mechanisms of action and developing cancer models that can more accurately represent tumor physiology. Survivin and telomerase reverse transcriptase (TERT), two important cancer biomarkers overexpressed in 85% of tumor tissues, play crucial roles in anti-apoptosis and unlimited cell proliferation in carcinogenesis, as well as drug resistance in cancer therapy. In this research, we have developed survivin and TERT reporter gene assays for high-throughput screening (HTS) of drugs downregulating these genes as anticancer therapeutics. Cancer cells were engineered to express fluorescent proteins controlled by TERT and survivin promoters, respectively, and were cultured in 3-dimensional (3D) scaffolds in microbioreactors to investigate drug effects by monitoring changes in culture fluorescence in real time. The novel 3-D assays offered molecular and physiological predictions of gene-specific responses to drugs, thus facilitating cancer drug discovery and development. Using survivin as the target, a dual-color fluorescent reporter gene assay performed in 3-D culture was developed. To demonstrate the drug's specific effect on the survivin promoter, MCF-7 breast cancer cells were engineered to express enhanced green fluorescent protein (EGFP) driven by a survivin promoter. The cell also expressed red fluorescent protein (RFP) under the control of a cytomegalovirus promoter as an internal control and to indicate cell number. The specific survivin expression was assessed by the green fluorescence normalized to the red fluorescence, which was used as an indicator to assess drug’s regulation on survivin. The internal control compensated for the variations in cell number, cell culture environment, fluorescent protein property, detection setting, and even for the reading interference from tested compounds, resulting in a more accurate evaluation of drug effect. The assay correctly detected the downregulation and upregulation of several anticancer compounds and demonstrated its reliability for high-throughput screening of survivin-targeted drugs. In this work, a 3-D cancer cell-based hTERT promoter assay was also developed for anticancer drug screening. Different lengths of the hTERT promoter were cloned with EGFP as reporting cassettes and were introduced in multiple cancer cell lines. The selected MCF7 reporter cell was able to correctly respond to the known hTERT inhibitors, pristimerin, epigallocatechin gallate and n-butylidenephthalide. The reporter cells were cultured on 3-D scaffolds to resemble in vivo tumor physiology and present in vivo like responses. The 3-D hTERT promoter assay was verified by pristimerin, and subsequently, other five anticancer compounds, including doxorubicin, cisplatin, paclitaxel, blasticidin, and tamoxifen, were tested for their effects on hTERT. The developed assay achieved an overall accuracy rate of over 80%, which demonstrated the feasibility of using 3D culture and EGFP as a reporter to screen hTERT inhibitors. In addition to the development of cancer cell-based assays, hTERT as a pluripotency marker to predict embryotoxic compounds during tissue development was also evaluated. The hTERT-EGFP reporting cassettes were introduced into mouse embryonic stem cells D3. The reportability was assessed by the corelated expression changes of EGFP and endogenous TERT during neural differentiation and after receiving toxicant stimuli. Our results showed that EGFP was able to indicate drug effect on human TERT expression level during ESC differentiation, which, however, was not always consistent with internal mouse TERT (mTERT) due to species difference. The results confirmed that drug effects on TERT were species-specific, especially at the beginning of differentiation. In conclusion, hTERT in mESCs reacted to embryotoxic compounds and the engineered mESC-hTERT-EGFP cells could be used for human teratogen prediction.
Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 814
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Book Description
Author: Matt Carter
Publisher: Academic Press
ISBN: 0323915612
Category : Medical
Languages : en
Pages : 416
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Book Description
Modern neuroscience research is inherently multidisciplinary, with a wide variety of cutting edge new techniques to explore multiple levels of investigation. This Third Edition of Guide to Research Techniques in Neuroscience provides a comprehensive overview of classical and cutting edge methods including their utility, limitations, and how data are presented in the literature. This book can be used as an introduction to neuroscience techniques for anyone new to the field or as a reference for any neuroscientist while reading papers or attending talks. - Nearly 200 updated full-color illustrations to clearly convey the theory and practice of neuroscience methods - Expands on techniques from previous editions and covers many new techniques including in vivo calcium imaging, fiber photometry, RNA-Seq, brain spheroids, CRISPR-Cas9 genome editing, and more - Clear, straightforward explanations of each technique for anyone new to the field - A broad scope of methods, from noninvasive brain imaging in human subjects, to electrophysiology in animal models, to recombinant DNA technology in test tubes, to transfection of neurons in cell culture - Detailed recommendations on where to find protocols and other resources for specific techniques - "Walk-through" boxes that guide readers through experiments step-by-step
Author: Paul L. Bartel
Publisher: Oxford University Press, USA
ISBN: 9780195109382
Category : Carrier proteins
Languages : en
Pages : 362
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Book Description
This volume, part of the Advances in Molecular Biology series, presents work by pioneers in the field and is the first publication devoted solely to the yeast two-hybrid system. It includes detailed protocols, practical advice on troubleshooting, and suggestions for future development. In addition, it illustrates how to construct an activation domain hybrid library, how to identify mutations that disrupt an interaction, and how to use the system in mammalian cells. Many of the contributors have developed new applications and variations of the technique.
Author: Nicola Casali
Publisher: Humana Press
ISBN: 9781617373916
Category : Science
Languages : en
Pages : 316
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Book Description
A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.
