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At least 289 records · Page 16Linked to original sources
PubMed2002

Cryptic plasmids of Mycobacterium avium: Tn552 to the rescue.

Plasmids have been described in almost all bacterial species analysed and have proven to be essential genetic tools. In many bacteria these extrachromosomal DNAs are cryptic with no known markers or function, which makes their characterization and genetic exploitation extremely difficult. Here we describe a system that will allow the rescue of any circular DNA (plasmid or phage) using an in vitro transposition system to deliver both a selectable marker (kanamycin) and an Escherichia coli plasmid origin of replication. In this study, we demonstrate the rescue of four cryptic plasmids from the opportunistic pathogen Mycobacterium avium. To evaluate the host range of the rescued plasmids, we have examined their ability to be propagated in Mycobacterium smegmatis and Mycobacterium bovis BCG, and their compatibility with other mycobacterial plasmids. In addition, we use a library of transposon insertions to sequence one plasmid, pVT2, and to begin a genetic analysis of plasmid genes. Using this approach, we identified a putative conjugative relaxase, suggesting this myco-bacterial plasmid is transferable, and three genes required for plasmid establishment and replication.

Base Sequence↗
PubMed2000

A new world view of genetics service models.

Identification of the components of the Human Genome and their relevance to health and disease is revolutionizing the provision of genetic services and all areas of health care. New genetic tools to diagnose, manage, and treat common diseases, along with Web-based innovations are changing the shape of how genetic services will be accessed and delivered. These advances represent a "bionic convergence" that will fundamentally transform medicine during the next few decades. The convergence of biotechnology and electronics is creating an expanding array of health care opportunities for clients and offers innovative opportunities for health promotion, restoration and management. Families and communities will soon be able to participate more fully in the direction and design of their own genetic health. Nurses with their long history of providing holistic, family-centered care in all practice settings, can help to create new dimensions to their practice to support their clients as they meet these health care innovations. This article explores a New World view of genetics services, and describes futuristic models for their provision. Nursing participation in and preparation for future genetics services also is described.

Ethics, Clinical↗
PubMed1996

Genetic differences between nitrate-negative and nitrate-positive C. pseudotuberculosis strains using restriction fragment length polymorphisms.

Corynebacterium pseudotuberculosis has been classified into two biotypes according to ability to breakdown nitrate (Biberstein et al., 1971). Restriction enzyme analysis (REA) has shown to reflect this differentiation, but numerous bands generated by this technique make interpretation difficult (Songer et al., 1988). Restriction fragment length polymorphism's (RFLP's) has become an accepted genetic tool and was used in this study to determine if differences in nitrate reduction and other phenotypic characteristics could be identified genetically. Thirteen C. pseudotuberculosis isolates from four species of domestic animals from different parts of the world were investigated for phenotypic and genetic differences. Three closely related bacteria, Corynebacterium ulcerans, Actinomyces pyogenes (previously C. pyogenes),and Rhodococcus equi (previously C. equi) were included in the study to determine if the RFLP bands were unique to C. pseudotuberculosis. All C. pseudotuberculosis isolates were positive for urease production. Some differences in maltose and sucrose fermentation ability and nitrate reduction were recorded. Genetic differences were identified between the nitrate-positive group and the nitrate-negative group using non-radioactive ribosomal RNA (rRNA) probes Southern blotted to restriction digests of ApaI, PstI, and SstI. A small number of bands were seen, with distinct differences between the nitrate-positive and the nitrate-negative strains. No genetic variations were seen between strains which reflected differences in carbohydrate fermentation. Strains isolated from different animal species and from different parts of the world could not be differentiated genetically using these three restriction enzymes.

Animals↗
PubMed2006

Protein kinase C alpha but not PKCzeta suppresses intestinal tumor formation in ApcMin/+ mice.

Members of the protein kinase C (PKC) family of serine/threonine kinases play key regulatory roles in numerous cellular processes, including differentiation and proliferation. Of the 11 mammalian PKC isoforms known, several have been implicated in tumor development and progression. However, in most cases, isotype specificity is poorly defined, and even contrary functions for a single PKC have been reported mostly because appropriate molecular and genetic tools were missing to specifically assess the contribution of single PKC isoforms in vivo. In this report, we therefore used PKC genetic targeting to study the role of PKCalpha and PKCzeta in colorectal cancer. Both isoforms were found to be strongly down-regulated in intestinal tumors of ApcMin/+ mice. A deletion of PKCzeta did not affect tumorigenesis in this animal model. In contrast, PKCalpha-deficient ApcMin/+ mice developed more aggressive tumors and died significantly earlier than their PKCalpha-proficient littermates. Even without an additional Apc mutation, PKCalpha knockout mice showed an elevated tendency to develop spontaneous intestinal tumors. Transcriptional profiling revealed a role for this kinase in regulating epidermal growth factor receptor (EGFR) signaling and proposed a synergistic mechanism for EGFR/activator protein and WNT/APC pathways in mediating intestinal tumor development.

Animals↗
PubMed2003

The genes and brains of mice and men.

The elucidation of the human genome presents a challenge for psychiatry-determining the impact of thousands of genes on brain functions relevant to mental disorders. For both historical and practical reasons, the mouse has become the mammal of choice for applying molecular genetic approaches to gene function. A working draft of the mouse genome has led to estimates that a mouse version may be identified for 99% of human genes. In accord with their genomic homologies, humans and mice share numerous features of brain organization and behavioral responses to many pharmacological agents. Technologies enabling the precise experimental manipulation of the mouse genome provide unprecedented opportunities for exploring genetic contributions to the regulation of complex behavior and to the pathophysiology and treatment of psychiatric disease. The formidable array of mouse molecular genetic tools are applied for two general strategies: 1) exploring the function of particular genes by generating lines of mice with precise genetic alterations and 2) searching broadly for those genes that regulate a particular biological trait of interest. Essential to the effective use of these technologies is the implementation of sound strategies for discerning the impact of genetic manipulations on mouse behaviors relevant to psychiatric conditions. These approaches are having a major impact-examples relevant to psychiatric disorders are discussed. However, advances in implementing and interpreting behavioral assays have not kept pace with molecular genetic technologies. To maximize the extent to which the revolution in mammalian genetics may be effectively applied to psychiatric research, new technologies and strategies for mouse behavioral assessment must be developed.

Animals↗
PubMedDate not supplied

Dissecting hypertension: the role of the 'new genetics'.

The tools of molecular genetics have recently been applied to hypertension, a common multifactorial disorder, with some success. Glucocorticoid-suppressible hyperaldosteronism, an inherited form of human hypertension due to the dominant inheritance of a chimaeric steroid 11 beta-hydroxylase/aldosterone synthase gene, has given an insight into the possible genetic factors involved in essential hypertension. Study of the aldosterone synthase and steroid 11 beta-hydroxylase genes has shown the presence of polymorphisms in both of these genes in human subjects; further studies may demonstrate genetic mutations with pathophysiological effects in patients with essential hypertension.

Cytochrome P-450 CYP11B2↗
PubMed2005

Plant genome analysis: the state of the art.

Plants are the basis for the survival of all "higher" organisms on Earth. Development of molecular genetics tools has allowed analysis of the structure, evolution, and function of whole plant genomes, rather than individual genes. DNA-based markers were instrumental in constructing detailed genetic maps of model plants and all major crop species. These molecular maps were the basis of physical maps and the first plant whole genome sequences. Comparative analysis based on genetic, cytogenetic, and physical maps and DNA sequence information provided new insights into the evolution of plant nuclear and organellar genomes. Mapping factors controlling Mendelian and quantitative traits made possible the cloning and functional characterization of novel genes, which function in plant development, adaptation to biotic and abiotic stress, or in the formation of other agronomic characters. The parallel analysis of all transcripts, proteins, and metabolites present in plant cells or tissues has generated information that may lead to a better integrated understanding of genome function. Postfunctional analysis of natural variation of gene function and its effects on phenotype is envisaged to provide new diagnostic and therapeutic molecular tools for applications in plant breeding, adaptation, and ecology.

DNA, Plant↗
PubMed2002

Immune modulation: the genetic approach.

As part of a comprehensive strategy to combat diseases, improving genetically resistance to diseases and therefore immune capacities of animals is more and more desirable. However, research is still needed to develop genetic tools that may be used. In this search, lines selected for various immune responses are used to study relevant immune markers. Chickens have been selected for six generations for three different in vivo immune responses: high antibody response, high cell-mediated immune response and high phagocytic activity. Each line, selected for one trait, showed significant increase in immune capacity for this trait. In addition, results showed clearly independence between the three immune responses analyzed, meaning that a global approach is needed to improve immune capacity. Selected lines allow to follow genetic markers linked to immune response genes. In the different lines, different patterns in MHC gene frequency were observed and MHC alleles differed in their effects according to the immune trait. Some correlations were found between immune responses and production traits. The selected lines will be used to find other "known" immune response genes or "anonymous" genetic markers, which may become the future tools to modulate immune responsiveness of animals.

Animals↗
PubMed2000

Nested chromosomal deletions induced with retroviral vectors in mice.

Chromosomal deletions, especially nested deletions, are major genetic tools in diploid organisms that facilitate the functional analysis of large chromosomal regions and allow the rapid localization of mutations to specific genetic intervals. In mice, well-characterized overlapping deletions are only available at a few chromosomal loci, partly due to drawbacks of existing methods. Here we exploit the random integration of a retrovirus to generate high-resolution sets of nested deletions around defined loci in embryonic stem (ES) cells, with sizes extending from a few kilobases to several megabases. This approach expands the application of Cre-loxP-based chromosome engineering because it not only allows the construction of hundreds of overlapping deletions, but also provides molecular entry points to regions based on the retroviral tags. Our approach can be extended to any region of the mouse genome.

Animals↗
PubMed2004

Genetic models in pathogenesis.

To decipher the complexity of host-pathogen interactions the widest possible range of model hosts and of analytical methods is required. As some virulence mechanisms and certain host responses have been conserved throughout evolution, even simple organisms can be used as model hosts to help our understanding of infectious diseases. The availability of molecular genetic tools and a cooperative community of researchers are pivotal to the emergence of model systems. In this review, we first summarize the genetic screens that can be used to identify pathogen virulence factors, then we present a comparative overview of existing or emerging genetically tractable host models.

Animals↗
PubMed2000

Peroxisome biogenesis in the yeast Yarrowia lipolytica.

Extensive peroxisome proliferation during growth on oleic acid, combined with the availability of excellent genetic tools, makes the dimorphic yeast, Yarrowia lipolytica, a powerful model system to study the molecular mechanisms involved in peroxisome biogenesis. A combined genetic, biochemical, and morphological approach has revealed that the endoplasmic reticulum (ER) plays an essential role in the assembly of functional peroxisomes in this yeast. The trafficking of some membrane proteins to the peroxisomes occurs via the ER, results in their glycosylation in the ER lumen, does not involve transit through the Golgi, and requires the products of the SEC238, SRP54, PEX1, and PEX6 genes. The authors' data suggest a model for protein import into peroxisomes via two subpopulations of ER-derived vesicles that are distinct from secretory vesicles. A kinetic analysis of the trafficking of peroxisomal proteins in vivo has demonstrated that membrane and matrix proteins are initially targeted to multiple vesicular precursors that represent intermediates in the assembly pathway of peroxisomes. The authors have also recently identified a novel cytosolic chaperone, Pex20p, that assists in the oligomerization of thiolase in the cytosol and promotes its targeting to the peroxisome. These data provide the first evidence that a chaperone-assisted folding and oligomerization of thiolase in the cytosol is required for the import of this protein into the peroxisomal matrix.

Fungal Proteins↗
PubMed2006

The use of genetic clines to estimate dispersal distances of marine larvae.

Many unresolved issues in the ecology and evolution of marine populations center on how far planktonic larvae disperse away from their parents. Genetic tools provide a promising way to define the spatial spread of larvae, yet their accurate interpretation depends on the extent to which genetic loci are under selection. Genetic clines, geographic zones in which genetically differentiated populations interbreed, provide opportunities to explicitly and simultaneously quantify the relative roles of selection and dispersal. Here, we review the theory and analysis of genetic clines and apply these techniques to published studies of multilocus clines in the sea. The geographic width of a stable genetic cline is determined by a balance between the homogenizing effects of dispersal and the diversifying effects of selection. For marine researchers, the power of genetic clines is that, if selection and clinal width are quantified, then the average geographic distances that larvae move can be inferred. Measuring selection or dispersal through laboratory or field-based experimentation is possible, though logistically difficult, for pelagically dispersed organisms. Instead, dispersal may be more robustly quantified from the degree of linkage disequilibrium between two or more loci, because linkage disequilibrium integrates selection across multiple life stages and generations. It is also relatively insensitive to whether exogenous or endogenous selection operates. Even without quantifying linkage disequilibrium, the theory of genetic clines indicates that the average dispersal distance of larvae is a fraction (i.e., generally <35%) of the clinal width. Because cline theory is based on several underlying assumptions, including near-equilibrium between selection and migration, the dispersal distances inferred from empirical data should be of the correct order but may not be precise. Even so, such estimates of larval dispersal are valuable, as they can be utilized to design appropriate scales for future investigations and provide some guidance to conservation efforts.

Animal Migration↗
PubMed1977

Microcell-mediated transfer of murine chromosomes into mouse, Chinese hamster, and human somatic cells.

In this report, we describe the production and characterization of proliferating hybrid cell populations generated by fusion of murine microcells with intact mouse, Chinese hamster, and human recipient cells. The microcell hybrids so produced contained one to five intact murine chromosomes derived from the microcell donor. these transferred chromosomes were maintained as functioning genetic elements in the hybrid cells. Our results firmly establish subnuclear particle-mediated chromosome transfer as a valid somatic cell genetic tool.

Animals↗
PubMed1987

Detection of novel centromeric polymorphisms associated with alpha satellite DNA from human chromosome 11.

The pericentromeric region of human chromosomes is composed of diverse classes of repetitive DNAs, which provide a rich source of genetic variability. Here, we describe two novel centromeric polymorphisms associated with a subset of alpha satellite repetitive DNA, D11Z1, which is specific for human chromosome 11. Segregation and inheritance of the polymorphisms are demonstrated and their relative frequencies are determined. These polymorphisms may be useful genetic tools for distinguishing between individual chromosome 11 centromeres. In addition, these polymorphisms may be applied to the development of a centromere-based genetic linkage map of chromosome 11. Molecular models for the generation of these polymorphisms are discussed.

Centromere↗
PubMed2000

It takes two transposons to tango: transposable-element-mediated chromosomal rearrangements.

Transposable elements (TEs) promote various chromosomal rearrangements more efficiently, and often more specifically, than other cellular processes(1-3). One explanation of such events is homologous recombination between multiple copies of a TE present in a genome. Although this does occur, strong evidence from a number of TE systems in bacteria, plants and animals suggests that another mechanism - alternative transposition - induces a large proportion of TE-associated chromosomal rearrangements. This paper reviews evidence for alternative transposition from a number of unrelated but structurally similar TEs. The similarities between alternative transposition and V(D)J recombination are also discussed, as is the use of alternative transposition as a genetic tool.

Animals↗
PubMed1996

Autonomous transposition of the tobacco retrotransposon Tto1 in rice.

The complete nucleotide sequence of the tobacco retrotransposon Tto1, one of the few active retrotransposons of plants, was determined. The sequence analysis suggests that Tto1 carries all functions required for autonomous transposition through reverse transcription. Gene organization and the nature of the transcription product suggest that Tto1 uses a gene expression mechanism different from those employed by retroviruses and most retrotransposons to regulate Gag and Pol stoichiometry. Tto1 was introduced into rice to study its autonomous transposition in heterologous hosts. Transcription and transposition of Tto1 were observed in rice cells. To probe the autonomous transposition through reverse transcription, a modified Tto1 retrotransposon in which part of a reverse transcriptase gene was replaced with an intron-containing hygromycin resistance gene was constructed and introduced into rice cells. Loss of the intron was observed only when intact Tto1 was cotransfected. These results indicate that Tto1 can transpose autonomously through reverse transcription and that the host factors required for transposition are conserved among monocots (class Magnoliopsida; rice) and dicots (class Liliopsida; tobacco), which diverged approximately 200 million years ago. These findings are discussed in relation to the regulation and evolution of retrotransposons and the possible use of Tto1 as a molecular genetic tool.

Base Sequence↗
PubMed2005

Development and validation of tools to assess genetic discrimination and genetically based racism.

It is possible that communication from mass media, public health or consumer advertising sources about human genetics and health may reify stereotypes of racialized social groups, perhaps cueing or exacerbating discriminatory and racist attitudes. This research used a multifaceted approach to assess lay perceptions of genetic discrimination and genetically based racism (N = 644). Two tools for use in strategic planning efforts associated with communicating about human genetics and health, the genetic discrimination instrument (GDI) and the genetically based racism instrument (GBRI), were derived. The GDI emerged as having five dimensions associated with lay perceptions of genetic discrimination. The GBRI was found to be unidimensional. Scale validation activities supported the tools' concurrent and discriminant validity characteristics. Significant differences between blacks and whites on the criminal control rights, social reproductive rights and employer rights factors as well as the GBRI were found. We recommend application of these screening tools prior to national dissemination of messages associated with genes and disease susceptibility, including school and university-based curricula.

Adult↗
PubMedDate not supplied

Identification of Candida albicans genes that induce Saccharomyces cerevisiae cell adhesion and morphogenesis.

Morphogenesis and adhesion to host tissues and medical devices contribute to the virulence of Candida albicans, the most common fungal pathogen isolated from humans. However, identification of molecular mechanisms of C. albicans adhesion and morphogenesis has been impaired by the lack of effective molecular and genetic tools available for this organism. Saccharomyces cerevisiae provides an attractive model system for studying C. albicans adhesion and morphogenesis because of its well-characterized genetics and gene expression systems. To gain insight into the genetic mechanisms of C. albicans adhesion and morphogenesis, we used a parallel plate flow chamber to screen and quantitatively characterize attachment to polystyrene of an adhesion-deficient nonfilamentous flo8Delta S. cerevisiae strain expressing a C. albicans genomic library. We identified six C. albicans genes that are capable of promoting cell adhesion and pseudohyphal development in S. cerevisiae. We also analyzed the ability of these adhesion-promoting genes to regulate the expression of FLO11, which encodes an endogenous S. cerevisiae adhesin. One C. albicans gene, EAP1, appears to directly mediate adhesion and morphogenesis while the remaining five (EAP2, SWI1, MSB1, AAF1, and TEC1) upregulate expression of endogenous S. cerevisiae adhesins. These results suggest that S. cerevisiae is a useful system for molecular characterization of factors that regulate C. albicans adhesion and morphogenesis and that parallel plate flow chamber-based adhesion assays can be used in conjunction with genetic screens to identify molecular mechanisms regulating fungal cell adhesion.

Biotechnology↗