Gene amplification in mammalian cells a comprehensive guide

Cover of: Gene amplification in mammalian cells |

Published by M. Dekker in New York .

Written in English

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Subjects:

  • Gene amplification.,
  • Gene amplification -- Research -- Methodology.

Edition Notes

Includes bibliographical references and index.

Book details

Statementedited by Rodney E. Kellems.
ContributionsKellems, Rodney E., 1947-
Classifications
LC ClassificationsQH450.3 .G46 1993
The Physical Object
Paginationxxvii, 543 p. :
Number of Pages543
ID Numbers
Open LibraryOL1722561M
ISBN 100824787560
LC Control Number92026048

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Buy Gene Amplification in Mammalian Cells: A Comprehensive Guide on FREE SHIPPING on qualified ordersCited by: Gene amplification in mammalian cells: strategies for protein production Kellems This could result in saving considerable time and mate- rial.

Cell lines in which amplification occurs at an unwork- ably low frequency may become amenable to gene trans- fer and amplification with the help of these by: Serves as a comprehensive review to the substantial impact of gene amplification in molecular biology, genetic engineering and medical science.

The book covers the mechanism of gene amplification, organization and structure of amplified genes. Reporter genes for monitoring gene expression in mammalian cells (J.L.

Cook). Gene transfer and amplification in mammalian cells (F. Wurm). Co-transfer of multiple plasmids/viruses to introduce several genes in mammalian cells (F. Wurm). Optimization of plasmid backbone for gene expression in mammalian cells (D. Scherman).

cultured mammalian cells, can be selected for gene am- plification is greater than standard mutation rates, sug- gesting that gene amplifications are relatively frequent. This review describes current studies of gene amplification in mammalian cells, with emphasis on the acquisition of.

Gene amplification is a phenomenon occurring frequently in immortalized mammalian cells Chromosome segments with plasmid sequences are excised and circularized to form replicating episomes, which CHO cells with highly amplified, chromosomally localized DNA sequences are Cited by: Gene amplification in mammalian cells was first described as a cellular phenomenon that counteracts the increasing dosage of clastogenic drugs.

Levels of the enzyme dihydrofolate reductase (DHFR) increase dramatically in mouse cells during stepwise selection with methotrexate (MTX), an inhibitor of. Gene Amplification in Somatic Mammalian Cells Amplification of a number of genes has been reported in somatic mammalian cells, including the CAD gene (Wahl et al., ), the metallothionein gene (Beach and Palmiter, ), the HGPRT gene (Brennard et al., ), hydroxymethylglutaryl CoA reductase (Chin et al., ), adenosine deaminase (Yeung et al., ), glutamine synthetase Cited by:   DNA amplification is rare in normal human cells.

Proc Natl Acad Sci U S A. Mar; 87 (5)– [PMC free article] Schimke RT. Gene amplification in cultured cells. J Biol Chem. May 5; (13)– Stark GR, Debatisse M, Giulotto E, Wahl GM. Recent progress in understanding mechanisms of mammalian DNA amplification. by: Gene amplification may be viewed as a different type of alteration in genome structure; it increases the number of copies of a gene within a cell.

Gene amplification results from repeated rounds of DNA replication, yielding multiple copies of a particular region (Figure ).

The amplified DNA sequences can be found either as free extrachromosomal molecules or as tandem arrays of sequences within a.

Melera, P.W. and Biedler, J.L. Molecular and cytogenetic analysis of multidrug resistance-associated gene amplification in Chinese hamster, mouse sarcoma, and human neuroblastoma cell. In Molecular and Cellular Biology of Multidrug Resistance in Tumor Cells, edited by I.B. Roninson. Plenum Press: New York, pp.

– Google ScholarCited by: In this system, the spontaneous rate of gene amplification has been determined to 10 −3 events per cell division (5). Amplification is enhanced not only by MTX treatment but also by other agents which affect nucleotide metabolism (3), or by treatment of cells with tumor promoters or mutagens (6, 7).Cited by: 6.

Cytogenetic consequences of gene amplification: HSRs and DMs in antifolate-resistant and multidrug-resistant Chinese hamster and mouse cells and in human neuroblastoma cells with amplified N-myc protooncogenes --Organization and structure of dihydrofolate reductase amplicons --Analysis of gene amplification by in-gel DNA renaturation --Molecular cytogenetic characterization of the sequential development of multidrug resistance in two panels of colchicine-resistant cell.

The key is that the EcoRI site is within the kan r gene, so when a piece of human DNA is inserted there, the gene's function is destroyed.

Figure Screening Clones All E. coli cells transformed by the vector, whether it carries human DNA or not, can grow in the presence of ampicillin. Gene amplification is much more likely to occur in tumor cells that have already acquired genetic instability through alterations in cell cycle checkpoint pathways.

Gene amplification can also occur as an artifact when tumor cells are cultured in vitro. HISTOLOGY. In conclusion, the IR/MAR gene amplification method is a novel and efficient platform for recombinant antibody production in mammalian cells, which rapidly and easily enables the establishment of.

Genre/Form: Electronic books: Additional Physical Format: Print version: Gene amplification in mammalian cells. New York: M. Dekker, © (DLC) Gene transfer and gene amplification in mammalian cells / Florian M. Wurm and Martin Jordan ; Ch. Co-transfer of multiple plasmids-viruses as an attractive method to introduce several genes in mammalian cells / Martin Jordan and Florian M.

Wurm ; Ch. Optimization of plasmid backbone for gene expression in mammalian cells /. Each volume of Advances in Pharmacology provides a rich collection of reviews on timely topics. Emphasis is placed on the molecular bases of drug action, both applied and experimental.

Vol GeneTherapy, features important new research on gene transfers and therapy in the herpes simplex virus, anti-tumor immunity, steroid receptors, cystic fibroses, and Features* It provides an. Free shipping for non-business customers when ordering books at De Gruyter Online. Please find Permanent Gene Expression in Mammalian Cells: Gene Transfer And Selection.

Aspects of Gene Transfer and Gene Amplification in Recombinant Mammalian Cells. Wurm, Florian Μ. Pages Get Access to Full Text. Isolation of. Gene amplification methods play a crucial role in establishment of cells that produce high levels of recombinant protein.

However, the stability of such cell lines and the level of recombinant Author: Takahito Ohira, Koichi Miyauchi, Narumi Uno, Narumi Uno, Noriaki Shimizu, Yasuhiro Kazuki, Mitsuo Os. Amplified genes in cancer cells reside on extrachromosomal double minutes (DMs) or chromosomal homogeneously staining regions (HSRs).

We used a plasmid bearing a mammalian replication initiation region to model gene amplification. Recombination junctions in the amplified region were comprehensively identified and by: Plasmids A Desktop Resource (1st Edition) 2 | P a g e Plasmids Introduction to Addgene’s Resource Any newcomer who joins a molecular biology lab will undoubtedly be asked to design, modify, or construct a plasmid.

Although the newcomer likely knows that a plasmid is a small circular piece of DNA found in bacterial cells, she mayFile Size: 2MB. conclusion, the IR/MAR gene amplification method is a novel and efficient platform for recombinant antibody production in mammalian cells, which rapidly and easily enables the establishment of.

The amplification mechanism of the DHFR gene in MTX-resistant cells has been frequently employed for recombinant protein production in mammalian cells (12, 13). Because the production of protein pharmaceuticals, including cytokines and humanized antibodies, requires the use of cultured mammalian cells, a robust method for the production of Cited by:   Gene amplification is a widely used strategy for making pharmaceutical proteins in mammalian cells.

Since our DRCR system shares features with gene amplification seen in CHO cells, we speculate that our system could produce as much recombinant protein as a conventional CHO cell expression by: The use of vectors based on gene-amplification for the expression of cloned genes in mammalian cells.

In DNA Cloning, Vol. III, ed. Glover. IRL Press, Oxford,pp. –Author: Mary M. Bendig. In conclusion, the IR/MAR gene amplification method is a novel and efficient platform for recombinant antibody production in mammalian cells, which rapidly and easily enables the establishment of stable high-producer cell by: This chapter discusses some aspects of gene transfer and amplification in dihydrofolate reductase-deficient (dhfr-deficient) Chinese Hamster Ovary (CHO) cells, one of the most popular mammalian.

Gene amplification is an experimental strategy for increasing protein production in mammalian cells. Co-amplification of the target gene by genetically linking it to one or more selectable and Author: Carlo Alberto Redi.

The production of proteins in mammalian cells is an important tool in numerous scientific and commercial areas. For example, proteins for human therapy, vaccination or diagnostic applications are typically produced in mammalian cells.

Gene cloning, protein engineering, biochemical and biophysical characterization of proteins also require the use of gene expression in mammalian cells. Escherichia coli aspartate transcarbamylase: a novel marker for studies of gene amplification and expression in mammalian by: 8.

Characterization of an episome produced in hamster cells that amplify a transfected CAD gene at high frequency: functional evidence for a mammalian replication origin.

Molecular and Cellular Biology. Cited by: Drug-selected intrachromosomal gene amplification by breakage-fusion-bridge (BFB) cycles is well documented in mammalian cells, but factors governing this mechanism are not clear. Here, we show that only some clastogenic drugs induce drug resistance through intrachromosomal amplification.

We strictly correlate triggering of BFB cycles to induction of fragile site by: Drug-selected intrachromosomal gene amplification the selected gene (Windle et al., ; Toledo et al., by breakage-fusion-bridge (BFB) cycles is well docb, ), called “double minutes” (DMs) (Spriggs umented in mammalian cells, but factors governing et al., ; Cowell, ) or “episomes” (Carroll et al, this mechanism are.

using mammalian cells for protein production, can be over-come by gene amplification in CHO cells. For CHO cells, powerful gene amplification systems, such as dihydrofolate reductase (DHFR)-mediated or glutamine synthetase (GS)-mediated gene amplification, are available.

Thirdly, CHO cells have the capacity for efficient post-translational mod-File Size: KB. A One-Step Gene Amplification System for Use in Cultured Mammalian Cells and Transgenic Animals Article (PDF Available) in Transgenic Research 10(2) May with Reads. Strategy for amplifying a gene of interest on the MAC.

(A) Amplification mechanism of IR/MAR plasmid in transfected cells suggested by our previous studies (5,7,10).The plasmid bearing IR/MAR (red arrow) is multimerized as a circular direct repeat in transfected cells that frequently recombines with DMs or any co-transfected DNA and can integrate into random sites in the Cited by: 1.

Cells that produce higher levels of DHFR protein, and the adjoining recombinant protein gene, can compensate for the more rapid turnover of the DHFR protein and survive the selection process. This effect can complement MTX amplification to reduce the amount of MTX and shorten the time needed to generate a high-expressing by: 6.

N6-methyladenine is incorporated into mammalian genome by DNA polymerase dATP with a ratio of was used for PCR amplification, of alkbh1 gene in HEKT cells.

This volume provides a broad, state-of-the-art coverage of diverse technical topics in gene expression in mammalian cells, including the development of vectors for production of proteins in cultured cells, in transgenic animals, vaccination, and gene therapy; progress in methods for the transfer of genes into mammalian cells and the optimization and monitoring of gene expression; advances in.Get this from a library!

Gene transfer and expression in mammalian cells. [Savvas C Makrides;] -- The production of proteins in mammalian cells is an important tool in numerous scientific and commercial areas.

For example, proteins for human therapy, vaccination or diagnostic applications are.

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