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Topic: Mutagenesis

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	Site-Directed Mutagenesis by Janet Mosley
		After performing the site-directed mutagenesis technique, the mutated double-stranded DNA can be taken and introduced back into the original cell from which the gene of interest was found to see what effect, if any, that the mutation has on the cell.
		One use of site-directed mutagenesis is modifying the structure of clinically useful proteins in order to either decrease unwanted side effects or to increase the effectiveness of the proteins.
		Through the use of site-directed mutagenesis, researchers have been able to produce a version of the thrombin molecule that loses nearly all of its procoagulant activity, yet allowing the molecule to retain its anticoagulant activity at the same time.
	www.samford.edu /~gekeller/mosley.html (838 words)

	Site-directed mutagenesis - Wikipedia, the free encyclopedia
		Site-directed mutagenesis is a molecular biology technique in which a mutation is created at a defined site in a DNA molecule, usually a circular molecule known as a plasmid.
		In general, site-directed mutagenesis requires that the wild-type gene sequence be known.
		An important milestone in modern molecular biology came in 1978 with the first description of site-directed mutagenesis using oligonucleotides for in vitro synthesis of mutant DNA.
	en.wikipedia.org /wiki/Site-directed_mutagenesis (422 words)

	NIH Guide: MOUSE MUTAGENESIS AND PHENOTYPING: NERVOUS SYSTEM AND BEHAVIOR
		Neuroscience-focused mutagenesis and phenotyping facilities established by this RFA are expected to serve as a national resource by producing a bank of mouse strains that harbor a wide range of mutations affecting murine nervous system function and behavior.
		Large-scale, genome-wide mutagenesis is a powerful approach for the analysis of gene function, and has historically played a major role in the evolution of our understanding of the biology of viruses, bacteria, fungi, the nematode, Caenorhabditis elegans, the fruit fly, Drosophila melanogaster, and most recently, the zebrafish, Danio rerio.
		While mutagenesis of multiple regions of varying size across the whole mouse genome will be conducted, it is expected that priorities for genomic regions and phenotypes of particular interest will be established by NSC.
	grants.nih.gov /grants/guide/rfa-files/RFA-MH-99-007.html (6929 words)

	PA-01-070: DEVELOPMENT OF ZEBRAFISH MUTAGENESIS AND SCREENING TOOLS
		While this large-scale screen focused primarily on developmental defects, it is clear that saturation mutagenesis of the zebrafish genome will be an indispensable means of achieving a complete understanding not only of development, but also of the physiology, behavior, aging, and disease processes of this vertebrate.
		Interests include, but are not limited to, development of new methods for mutagenesis and/or phenotypic characterization that would be of use in research on a wide range of diseases or organs, particularly if these methods could be applied to other animal models as well as the zebrafish.
		Biomaterials (e.g., pathogen-free mutant animals, preserved sperm and embryos, etc.) and other patentable research resources (e.g., genetic and phenotypic screens, mutagenesis protocols, and genetic and phenotypic data for all mutant strains, etc.) developed in projects funded as a result of this PA are expected to be made available and distributed to the broader scientific community.
	grants.nih.gov /grants/guide/pa-files/PA-01-070.html (4789 words)

	NIDA Genetics Workgroup: ENU MUTAGENESIS
		Harwell ENU Mutagenesis Program The purpose of the current mutagenesis programme is to generate and make available approximately 100 F1 progeny of mutagenised animals per week for genome-wide dominant phenotypic screens.
		Mouse Mutagenesis for Developmental Defects The goal of project III is to determine the function of genes on mouse Chromosome 11 by saturating the chromosome with recessive mutations
		Australian ENU Mutagenesis Center This project is producing and screening large libraries of mutagenized C57BL/6 or TCR transgenic mice to identify genes controlling the immune response.
	www.nida.nih.gov /about/organization/genetics/mouse/enu (438 words)

	[9G] Site-directed Mutagenesis (Site not responding. Last check: 2007-10-21)
		Site-directed mutagenesis has been widely used in the study of protein functions.
		This method was first developed by Michael Smith who was awarded a Nobel Prize in 1993 for this contribution.
		Site-directed mutagenesis can also be achieved by using PCR (more info).
	www.web-books.com /MoBio/Free/Ch9G.htm (186 words)

	Localized mutagenesis (Site not responding. Last check: 2007-10-21)
		Such localized mutagenesis is especially useful for obtaining rare point mutations in or near a gene of interest (for example, temperature-sensitive mutations in a gene, mutations in the promoter or operator of a gene, or mutations that affect amino acids at the active site of an enzyme).
		The extent of mutagenesis of the phage particles can be monitored indirectly by following decrease in phage titer (or "killing") due to mutations in essential phage genes, or directly by following the increase in clear plaque phage mutants in the lysate.
		In addition to mutagenesis of DNA in phage particles, this is also an effective method for isolating mutations in purified plasmid DNA.
	www.sci.sdsu.edu /~smaloy/MicrobialGenetics/topics/transposons/Local-mut.html (449 words)

	Environmental And Molecular Mutagenesis Journal - (Site not responding. Last check: 2007-10-21)
		Mutagenesis is an international multi-disciplinary journal designed to bring...
		Environmental and Molecular Mutagenesis, 44 (3): 189-189, 2004...
		The study of environmental mutagenesis is a multidisciplinary activity.
	molecular.fmqg.com /index.php?k=environmental-and-molecular-mutagenesis-journal (508 words)

	RFA-HD-00-004: MUTAGENESIS SCREENS/PHENOTYPING TOOLS FOR ZEBRAFISH
		Saturation mutagenesis of the zebrafish genome will be an indispensable means of achieving a complete understanding of the development, physiology, and behavior of this vertebrate.
		Biomaterials (pathogen-free mutant animals, preserved sperm and embryos) and other patentable research resources (e.g., genetic and phenotypic screens, mutagenesis protocols, and genetic and phenotypic data for all mutant strains) produced in projects funded by this RFA are expected to be made available and distributed to the broader scientific community.
		For this reason, NIH is concerned that patents on mutant fish, embryos, and sperm, genetic and phenotypic screens, mutagenesis protocols, and genetic and phenotypic data for all mutant strains and other research resources might have a chilling effect on the future development of products and information that may improve the public health.
	grants.nih.gov /grants/guide/rfa-files/RFA-HD-00-004.html (5757 words)

	Phenotype Mutagenesis (Site not responding. Last check: 2007-10-21)
		In contrast to genotype-driven mutagenesis, the phenotypic approach may still lag by a significant distance – basically, phenotype-based screens cannot be conducted in an efficient, large scale fashion (Schimenti, 1998).
		The technical route of undertaking a phenotype-driven mutagenesis is to modify the nucleotides by pairing the original nucleotide to an incorrect complement.
		In either case the identification of the mammalian Clock gene is expected to reveal new insights to the control of sleep and its disorders, jet lags and perhaps pathologies related to aging in humans.
	dragon.zoo.utoronto.ca /~J02T0202C/phenotype.html (1742 words)

	[No title] (Site not responding. Last check: 2007-10-21)
		ISBN 1-898486-01-8 1995 Horizon Scientific Press, PO Box 1, Wymondham, Norfolk, U.K. In vitro site-directed mutagenesis is an invaluable technique for studying protein structure-function relationships, gene expression and vector modification.
		Use of PCR in site-directed mutagenesis accomplishes strand separation by using a denaturing step to separate the complementing strands and allowing efficient polymerization of the PCR primers.
		First, it is often desirable to reduce the number of cycles during PCR when performing PCR-based site-directed mutagenesis to prevent clonal expansion of any (undesired) second-site mutations.
	wheat.pw.usda.gov /homepage/lazo/methods/lazo/pcr-mut.html (522 words)

	Directed Mutagenesis and Protein Engineering
		However, subtilisin is inactivated by bleach, and it was found that this inactivation was due to oxidation of the amino acid methionine at position 22 of the subtilisin molecule.
		One of the challenges of combinatorial mutagenesis is the selection of molecules with desired properties.
		This is particularly a problem if degeneracy has been introduced at a large number of codons simultaneously, and thus the expected frequency of molecules with desired properties may be very low.
	photoscience.la.asu.edu /photosyn/courses/BIO_343/lecture/protein.html (771 words)

	Mutagenesis -- Instructions to Authors
		Mutagenesis is an international multi-disciplinary journal designed to bring together research aimed at the identification, characterization and elucidation of the mechanisms of action of physical, chemical and biological agents capable of producing genetic change in living organisms and the study of the consequences of such changes.
		The conclusions of such studies are published in summary form in the Journal and copies of laboratory data for specific agents may be obtained directly from the Publisher, quoting Journal reference, test system and test agent.
		All manuscripts submitted to Mutagenesis are refereed for their pertinence, content and relevance to the scope of the journal.
	mutage.oxfordjournals.org /misc/ifora.dtl (2088 words)

	Mutagenesis of Neisseria meningitidis by In Vitro Transposition of Himar1 mariner -- Pelicic et al. 182 (19): 5391 -- ...
		Mutagenesis of Neisseria meningitidis by In Vitro Transposition of Himar1 mariner -- Pelicic et al.
		Mutagenesis of Neisseria meningitidis by In Vitro Transposition of Himar1 mariner
		In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation.
	jb.asm.org /cgi/content/full/182/19/5391 (4920 words)

	Mutagenesis methods
		Site-directed mutagenesis using a double-stranded DNA fragment as a PCR primer.
		Site-directed, recombination-mediated mutagenesis of a complex gene locus.
		The mutagenesis method is based on the use of single-stranded DNA and two primers, one mutagenic primer and a second correction primer whic...
	biowww.net /methods/browse-10.html (1005 words)

	Functional Genomics in the Mouse: Phenotype-Based Mutagenesis Screens -- Schimenti and Bucan 8 (7): 698 -- Genome ...
		The critical genetic reagent for region-specific mutagenesis is the chromosomal deletion.
		At present, mutagenesis of male germ cells with ENU is the most efficient method available for mutagenizing the mouse genome.
		Bode, V.C. Ethylnitrosourea mutagenesis and isolation of mutant alleles for specific genes located in the t region of mouse chromosome 17.
	www.genome.org /cgi/content/full/8/7/698 (6275 words)

	Site-saturation mutagenesis of the PALTAVETG motif in coxsackievirus A9 capsid protein VP1 reveals evidence of ...
		Site-saturation mutagenesis of the PALTAVETG motif in coxsackievirus A9 capsid protein VP1 reveals evidence of conservation of a periodic hydrophobicity profile -- Airaksinen et al.
		Site-saturation mutagenesis of the PALTAVETG motif in coxsackievirus A9 capsid protein VP1 reveals evidence of conservation of a periodic hydrophobicity profile
		Mutagenesis was performed on an M13 vector and the site-saturated insert was cloned back to the genome of CAV-9.
	vir.sgmjournals.org /cgi/content/full/80/8/1919 (4007 words)

	Mutagenesis,Custom Mutagenesis Services (Site not responding. Last check: 2007-10-21)
		Mutagenesis is performed by generating mutations in cloned segments of DNA.
		Bionexus custom mutagenesis services can produce mutations at an extremely high efficiency (approaching 100% in some cases).
		The experienced staff at Bionexus's Mutagenesis Services department can produce mutations with shortest turnaround time in 1-2 weeks and provide you with the purified plasmid DNA with the correct changes and.
	www.mutagenesisservices.net (273 words)

	mutagenesis
		A second round of mutagenesis can be performed on one of these lacZ+ tets mutants using the Ampicillin Knockout Oligonucleotide with the Tetracycline Repair Oligonucleotide for selection.
		By the alternate use of the two antibiotic resistances encoded by the vectors, it is possible to generate an indefinite number of mutations without subcloning the insert into another vector.
		This property is particularly useful for confirming the effect of a mutation by restoring the wild-type sequence through another round of mutagenesis.
	www.csun.edu /~hcbio027/biotechnology/lec5/mutagenesis.html (546 words)

	In vivo site-directed mutagenesis of Neurospora crassa
		In vivo site-directed mutagenesis of Neurospora crassa beta-tubulin gene by spheroplasts transformation with oligonucleotides
		In 1986 a mutant of the beta-tubulin gene of Neurospora crassa, resistant to the toxic compound benlate (Benomyl) was isolated (Orbach et al.
		Our data (Figure 1) show that while mutagenetic efficiency was directly proportional to the amount of the oligonucleotides used to at least 50 ug, there was an inverse correlation between the number of transformants and length of the oligonucleotide.
	www.fgsc.net /fgn43/callis.html (763 words)

	Bacterial Genetics - Site-directed Mutagenesis
		This approach to mutagenesis was pioneered by the late Michael Smith at the Univerity of British Columbia.
		He shared the 1993 Nobel Prize in Chemistry for his fundamental contributions to the establishment of oligonucleotide-based, site-directed mutagenesis and its development for protein studies.
		This is due to inefficiencies in synthesising the complementary DNA strand, primer annealing improperly to other sequences in template DNA, and to the host cell repair systems which recognise the heteroduplex and remove the mutation that you are attempting to introduce.
	www.mun.ca /biochem/courses/4103/topics/site-directed.html (753 words)

	Mutation Research - Genetic Toxicology and Environmental Mutagenesis. (Site not responding. Last check: 2007-10-21)
		Torres, A. Creus, R. Marcos, Genotoxic activity of four inhibitors of DNA topoisomerases in larval cells of Drosophila melanogaster as measured in the wing spot assay, Mutation Research - Genetic Toxicology and Environmental Mutagenesis 413 (2) (1998) pp.
		Holmalahti, J. Maki-Paakkanen, L. Kangas, A. von Wright, Genotoxicity of dihydroabikoviromycin, a secondary metabolite of Streptomyces anulatus, Mutation Research - Genetic Toxicology and Environmental Mutagenesis 368 (3-4) (1996) pp.
		K.K. Panda, J. Patra, B.B. Panda, Persistence of cadmium-induced adaptive response to genotixicity of maleic hydrazide and methyl mercuric chloride in root meristem cells of Allium cepa L.: Differential inhibition by cycloheximide and buthionine sulfoximine, Mutation Research - Genetic Toxicology and Environmental Mutagenesis 389 (2-3) (1997) pp.
	www.elsevier.com /cdweb/journals/13835718/viewer.htt?viewtype=keywords&rangeselected=82 (5939 words)

	Kits For Site-Directed Mutagenesis - Biocompare Technology Spotlight
		Transfection of E. coli with this DNA should produce wildtype and mutant phage.” The first experiments of this type involved the conversion of a trp codon to an amber codon, so there was an easy screen for the mutation.
		While these early mutagenesis experiments were groundbreaking, they weren’t very effective.
		Site-directed mutations with >80% mutagenesis efficiency DNA isolated from any source can be used, with no need for specialized vectors, host strains, or restriction sites.
	www.biocompare.com /spotlight.asp?id=145 (526 words)

	Moving forward with chemical mutagenesis in the mouse -- O'Brien and Frankel 554 (1): 13 -- The Journal of Physiology ...
		Toward the yeastification of mouse genetics: chemical mutagenesis of embryonic stem cells.
		N-ethyl-N-nitrosourea mutagenesis of a 6- to 11-cM subregion of the Fah-Hbb interval of mouse chromosome 7: Completed testing of 4557 gametes and deletion mapping and complementation analysis of 31 mutations.
		Behavior and mutagenesis screens: the importance of baseline analysis of inbred strains.
	jp.physoc.org /cgi/content/full/554/1/13 (4554 words)

	Eurofins Medigenomix GmbH: In vitro Mutagenesis
		By outsourcing in vitro mutagenesis projects to Medigenomix you can profit from an extremely short turnaround time even in demanding mutagenesis projects.
		The proven expertise of Medigenomix´ team in the field of in vitro mutagenesis and its optimized technology allows the delivery of mutated and verified vectors (purified DNA and bacterial clones) within a few days, thereby saving scientist´s valuable time.
		Site directed mutagenesis is a very powerful tool for studying protein structure-function relationships.
	www.medigenomix.de /en/molbio2E.html (251 words)

	Site-Directed Mutagenesis (Site not responding. Last check: 2007-10-21)
		The mutagenesis continues by the denaturing of the DNA a second time.
		After many cycles (about 40) there would be a sufficient number of mutated DNA strands to consider the non-mutated strands to be negligible.
		The mutagenesis is stopped, and the mutated section of DNA is then added to, or spliced, into the DNA of the bacteria (or other organism).
	www.che.utoledo.edu /nadarajah/webpages/mutagene.htm (572 words)

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