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Fluorescent In Situ Hybridization to Detect Transgene Integration into Plant Genomes

Fluorescent chromosome analysis technologies have advanced our understanding of genome organization during the last 30 years and have enabled the investigation of DNA organization and structure as well as the evolution of chromosomes. Fluorescent chromosome staining allows ...

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Establishing Substantial Equivalence: Transcriptomics

Regulatory authorities in Western Europe require transgenic crops to be substantially equivalent to conventionally bred forms if they are to be approved for commercial production. One way to establish substantial equivalence is to compare the transcript profiles of developing g ...

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Establishing Substantial Equivalence: Proteomics

Wheat is a major crop in world agriculture and is consumed after processing into a range of food products. It is therefore of great importance to determine the consequences (intended and unintended) of transgenesis in wheat and whether genetically modified lines are substantially equiva ...

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Establishing Substantial Equivalence: Metabolomics

Modern ‘metabolomic’ methods allow us to compare levels of many structurally diverse compounds in an automated fashion across a large number of samples. This technology is ideally suited to screening of populations of plants, including trials where the aim is the determination of uninte ...

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Design and Management of Field Trials of Transgenic Cereals

The development of gene transformation systems has allowed the introgression of alien genes into plant genomes, thus providing a mechanism for broadening the genetic resources available to plant breeders. The design and the management of field trials vary according to the purpose for wh ...

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Transgenic Wheat, Barley and Oats: Future Prospects

Following the success of transgenic maize and rice, methods have now been developed for the efficient introduction of genes into wheat, barley and oats. This review summarizes the present position in relation to these three species, and also uses information from field trial databases and the ...

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GM Risk Assessment

GM risk assessments play an important role in the decision-making process surrounding the regulation, notification and permission to handle Genetically Modified Organisms (GMOs). Ultimately the role of a GM risk assessment will be to ensure the safe handling and containment of the GMO; a ...

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Pronuclear Microinjection of Purified Artificial Chromosomes for Generation of Transgenic Mice: Pick-and-Inject Technique

Mammalian artificial chromosomes (MACs) are appealing vectors for transgenesis and gene therapy applications. Their principal benefits include large DNA-carrying capacity and the ability to replicate in synchrony with the host genome. In addition, introducing genes into art ...

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Modification of Human Bacterial Artificial Chromosome Clones for Functional Studies and Therapeutic Applications

With the completion of the sequencing phase of the Human Genome Project and the availability of fully sequenced clones containing intact functional loci from the various high-quality human P1- and F-plasmid derived/bacterial artificial chromosome (PAC/BAC) genomic libraries (1 ...

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Methodologies Applied to Domestic Animal Chromosomes

Chromosomes of domestic animals have attracted the attention of both scientists and breeders because chromosomal abnormalities have been strictly correlated with the reduced fertility in cattle carrying rob(1;29) (1). Domestic animal cytogenetics has expanded noticeably, e ...

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From Natural to Artificial Chromosomes: An Overview

The rationale for building artificial chromosomes (ACs) has been critically reviewed by various authors at different stages of this line of investigation, initiated in the early 1980s.

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Mapping of the Hotspots of Recombination in Human DNA Cloned as Yeast Artificial Chromosomes

Whole genome sequencing of several species, including our own, constitutes a good starting point for the study of various functions of the chromosome on a whole genome scale. Mapping of meiotic recombination hotspots (the sites in which recombination occurs more frequently than at an aver ...

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Genomic Imprinting and Its Effects on Genes and Chromosomes in Mammals

Genomic imprinting is a regulatory mechanism by which, at certain gene loci, one of the two alleles becomes repressed according to its parental origin. This chapter focuses on mammalian imprinting, which regulates not only the parental allele-specific gene expression on autosomal chr ...

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Centromeres and Neocentromeres

A functional centromere is formed by a chromosomal domain that very often, but not always, is recognizable by a primary constriction in metaphasic chromosomes. It is associated with a kinetochore through which a link is established with the microtubules, which pull the sister chromatids to ...

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Identification of Start Sites of Bi-Directional DNA Synthesis at Eukaryotic DNA Replication Origins

DNA replication is the key process for the conservation and transmission of genetic information in all living organisms. The bi-directional DNA replication does not start randomly along the genome but rather at precise chromosomal regions, defined as origins of DNA replication where, a ...

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Telomere Length Analysis and In Vitro Telomerase Assay

Barring exceptional instances, the DNA contained in eukaryotic chromosomes is linear. Linearity of the chromosomal DNA and the compartmentalized architecture of the eukaryotic cell are the two principle features that distinguish the prokaryotes from the eukaryotes and that have ...

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Telomerization of Mammalian Cells and Transplantation of Telomerized Cells in Immunodeficient Mice

Telomerization is an operational term for appparent immortalization sustained by prevention of telomere shortening. It is always necessary to use the term apparent because immortalization, if taken literally, requires one to demonstrate that cells divide forever, a property that ...

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Chromosome-Based Vectors for Mammalian Cells: An Overview

The functional organization of the eukaryotic chromosome was first elucidated at a molecular level in the budding yeast, Saccharomyces cerevisiae, providing the basis for the successful creation of yeast artificial chromosomes (YACs) (1). The structures that confer chromosome f ...

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Direct Cloning of Full-Length Cell Differentially Expressed Genes by Multiple Rounds of Subtractive Hybridization Based on Long-Distance PCR and Magne

Subtractive cloning is an ideal technique for identifying genes differentially expressed in two nuclear acids population (1). The polymerase chain reaction (PCR)-based subtraction is the method of choice when the starting samples are heterogeneous or difficult to obtain, which of ...

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Computer Programs for PCR Primer Design and Analysis

Heat will separate or &quote;melt&quote; double-stranded DNA into single-stranded DNA by disrupting its hydrogen bonds. T m (melting temperature) is the temperature at which half the DNA strands are single-stranded and half are double-stranded. T m characterizes the stability of the DNA h ...

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