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At least 325 records · Page 18Linked to original sources
PubMed1985

Genetic characterization of pyridine nucleotide uptake mutants of Salmonella typhimurium.

Two classes of pyridine nucleotide uptake mutants isolated previously in a strain of Salmonella typhimurium defective in both de novo NAD biosynthesis (nad) and pyridine nucleotide recycling (pncA) were analysed in terms of their genetic relationship to each other and their roles in the transport of nicotinamide mononucleotide as a precursor to NAD. The first class of uptake mutants, pnuA (99 units), failed to grow on nicotinamide mononucleotide (NMN) as a precursor for NAD. The second class, pnuB, grew on lower than normal levels of NMN and suppressed pnuA mutations. A third class of uptake mutant, pnuC, isolated in a nadB pncA pnuB background, also failed to grow on NMN. Transport studies and enzyme analyses confirmed these strains as defective in NMN uptake. A fourth locus, designated pnuD, was found to diminish NMN utilization in a nad pncA+ background. Tn10 insertions near pnuA, pnuC and pnuD were isolated and utilized in mapping studies. pnuA was found to map between thr and serB near trpR. The pnuC locus was cotransducible with nadA at 17 units while pnuD mapped at approximately 60 units. The biochemical and genetic data suggest that the pnuA and pnuC gene products cooperate in the utilization of NMN under normal conditions. A pnuB mutant, however, does not require the pnuA gene product for NMN uptake but does rely on the pnuC product. Fusion studies indicate that pnuC is regulated by internal NAD concentrations.

Biological Transport↗
PubMed1989

Use of promoter fusions in Drosophila genetics: characterization of a YP1-ADH fusion gene.

In Drosophila melanogaster the yolk protein (YP) genes are normally expressed only in the fat body and follicular epithelium of adult females--never in males or in larvae. We describe here a first step toward a genetic examination of the developmental controls that restrict the activity of the YP genes to adult female tissues. A YP1 promoter that contains the tissue-, temporal-, and sex-specific controlling elements for expression was fused to the reporter gene, alcohol dehydrogenase (Adh). The gene fusion was transformed into an Adh-deficient genotype. As assayed by a number of criteria, that the fusion gene is expressed in the same physiological manner as the endogenous yolk protein genes. The fusion gene's activity is modulated in trans by a temperature-sensitive allele of the sex determination gene, tra-2. The Adh enzyme serves as a selectable marker and therefore these flies are suitable for use in genetic screens for trans-acting mutations that affect the expression of the yolk protein genes.

Alcohol Dehydrogenase↗
PubMed2005

Biochemical and genetic characterization of Hmi1p, a yeast DNA helicase involved in the maintenance of mitochondrial DNA.

The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maintenance of the mitochondrial DNA (mtDNA) genome of Saccharomyces cerevisiae. Identified based on its homology with E. coli uvrD, the HMI1 gene product, Hmi1p, has been presumed to be involved in the replication of the 80 kb linear S. cerevisiae mtDNA genome. Here we report the purification of Hmi1p to apparent homogeneity and provide a characterization of the helicase reaction and the ATPase reaction with regard to NTP preference, divalent cation preference and the stimulatory effects of different nucleic acids on Hmi1p-catalysed ATPase activity. Genetic complementation assays indicate that mitochondrial localization of Hmi1p is essential for its role in mtDNA metabolism. The helicase activity, however, is not essential. Point mutants that lack ATPase/helicase activity partially complement a strain lacking Hmi1p. We suggest several possible roles for Hmi1p in mtDNA metabolism.

Adenosine Triphosphatases↗
PubMed2004

Genetic characterization of orf viruses isolated from various ruminant species of a zoo.

In the present study, an outbreak of proliferative dermatitis in musk ox (Ovibos moschatus), Sichuan takin (Budorcas taxicolor tibetana) and domestic Shetland sheep (Ovis aries) in a zoo is described. Skin lesions consisted of severe, persistent, multifocal, proliferative dermatitis in musk ox, and mild, transient, focal, dermatitis in the Sichuan takin and Shetland sheep. Parapoxviruses were isolated from skin lesions, and characterized by restriction enzyme analysis and partial gene sequencing. The results of this investigation indicate that the outbreak of proliferative dermatitis was due to infection by a single parapoxvirus, which is genetically closely related to other orf virus (ORFV) strains but distant to bovine papular stomatitis virus (BPSV) and pseudocowpox virus (PCPV).

Amino Acid Sequence↗
PubMed1995

Genetic characterization of small ovaries, a gene required in the soma for the development of the Drosophila ovary and the female germline.

The small ovary gene (sov) is required for the development of the Drosophila ovary. Six EMS-induced recessive alleles have been identified. Hypomorphic alleles are female sterile and have no effect on male fertility, whereas more severe mutations result in lethality. The female-sterile alleles produce a range of mutant phenotypes that affect the differentiation of both somatic and germline tissues. These mutations generally produce small ovaries that contain few egg cysts and disorganized ovarioles, and in the most extreme case no ovarian tissue is present. The mutant egg cysts that develop have aberrant morphology, including abnormal numbers of nurse cells and patches of necrotic cells. We demonstrate that sov gene expression is not required in the germline for the development of functional egg cysts. This indicates that the sov function is somatic dependent. We present evidence using loss-of-function and constitutive forms of the somatic sex regulatory genes that sov activity is essential for the development of the somatic ovary regardless of the chromosomal sex of the fly. In addition, the genetic mapping of the sov locus is presented, including the characterization of two lethal sov alleles and complementation mapping with existing rearrangements.

Alleles↗
PubMed1983

Antigenic and genetic characterization of H1 influenza viruses isolated from feral ducks and swine in Japan.

The strains of H 1 N 4 influenza A virus isolated from feral ducks in Japan in 1977-78 were compared to swine-origin H 1 N 1 viruses antigenically and genetically. Homologous characteristics were found among the H 1 N 4 isolates from feral ducks in hemagglutination-inhibition (HI) tests, viral RNA patterns on polyacrylamide gel electrophoresis and oligonucleotide mapping. Although the hemagglutinins of duck-origin viruses employed in this study were identified as H 1, the viruses were distinguishable from A/New Jersey/8/76 (H 1 N 1), A/duck/Alberta/35/76 (H 1 N 1) and the virus isolated from swine in Japan in the cross HI test. Also, the viral RNA patterns of the duck- and swine-origin H 1 viruses were found to be quite different, indicating that genetic reassortment of HA genes between them is unlikely. After H 1 N 4 virus of duck-origin was intranasally inoculated into pigs, a brief period of virus recovery with no serological response was observed; whereas swine-origin H 1 N 1 virus produced seroconversion in the pigs inoculated.

Animals↗
PubMed2001

Isolation and genetic characterization of two G3P5A[3] canine rotavirus strains in Italy.

Two strains of canine rotavirus were isolated from pups with clinical signs of gastroenteritis. Both strains were identified by polymerase chain reaction (PCR) as G3P5A[3], although restriction endonuclease analysis of the PCR amplicons revealed a genetic difference between the two isolates in the VP7 gene. The isolation in Italy of canine rotaviruses displaying the same VP7 and VP4 specificities as in the USA and in Japan, suggests that the G3 and P5A[3] types are highly conserved among canine rotavirus strains.

Animals↗
PubMed1997

Genetic characterization of populations of a de novo arisen sugar beet pest, Aubeonymus mariaefranciscae (Coleoptera, Curculionidae), by RAPD analysis.

The weevil Aubeonymus mariaefranciscae is an important pest of sugar beet crops in southern Spain that was first described as a new species in 1979. We have studied the genetic variability of this insect by RAPD analysis of 122 individuals using eight primers. The high resolution provided by random amplified polymorphic DNA (RAPDs), in combination with efficient genetic distance estimators, allowed a very detailed description of the ecology and evolution of the populations of this insect in the south of Spain. The results are compatible with a unique event of colonization, followed by the spreading of the weevil across the surrounding areas. A comparison of the results obtained with different genetic distance estimators is presented. RAPD is shown to be a powerful technique for reconstructing the phylogenetic history of insect populations, even if they have diverged recently, which seems to be the case for A. mariaefranciscae.

Animals↗
PubMed2002

Gene therapy for pancreatic cancer based on genetic characterization of the disease.

In order to develop an effective therapeutic intervention for patients with pancreatic cancer, we analyzed the association of loss of heterozygosity (LOH) with the clinicopathological features of the disease. Based on these results, we are developing a new gene therapy that targets the genetic character of pancreatic cancer, using mutant adenoviruses that are selectively replication-competent in tumor cells. LOH of 30% or more was observed on chromosome arms 17p (47%), 9p (45%), 18q (43%), 12q (34%), and 6q (30%). LOH of 12q, 17p, and 18q showed significant association with poor prognosis. These data strongly suggest that mutations of the putative tumor suppressor genes, TP53 and SMAD4, play significant roles in the disease progression. Based on this rationale, we are developing a new gene therapy that targets tumors without normal TP53 function. The E1B-55kDa-deleted adenovirus can selectively replicate in TP53-deficient human tumor cells, but not in cells with functional TP53. We evaluated the therapeutic effect of this E1B-55kDa-deleted mutant adenovirus on pancreatic cancer without normal TP53 function. The growth of a human pancreatic tumor in a severe combined immunodeficiency (SCID) mouse model was markedly inhibited by consecutive injections of the E1B-55kDa-deleted adenovirus. Furthermore, the replication-competent adenovirus is not only a strong weapon itself but it is also a useful carrier of genes that possess antitumor activities, as a virus vector specific to tumors without normal TP53 function.

Adenoviridae↗
PubMed1988

Genetic characterization and use of a restriction fragment length variant in the hypotrichous ciliate Euplotes crassus.

Two forms of a macronuclear DNA molecule differing in the presence or absence of a restriction endonuclease recognition site have been detected in the hypotrichous ciliate Euplotes crassus. Through a series of genetic crosses the two forms were shown to be allelic, being derived from a single micronuclear genetic locus. This restriction fragment length variant (RFLV) was used as a genetic marker to determine that the migratory and stationary pronuclei generated during mating can be genetically non-identical. In addition, the RFLV was used to investigate the efficiency of processing of the alternate alleles during macronuclear development and their subsequent transmission during vegetative growth. Little or no bias in the processing and/or amplification of the two alleles was observed during macronuclear development. During vegetative growth, however, changes in the relative amounts of the two alleles were observed.

Alleles↗
PubMed1992

Serologic and genetic characterization of Plasmodium vivax from whole blood-impregnated filter paper discs.

The presence in the New World of a variant strain of Plasmodium vivax (VK247) containing a unique circumsporozoite (CS) repeat domain was determined by the detection of antibodies to the variant CS protein and by genetic analysis of the CS gene from field isolates. Whole blood specimens were collected on filter paper from patients infected with P. vivax in Mexico and Peru. Plasmodium vivax DNA was eluted from filter paper samples and the CS gene was amplified by the polymerase chain reaction (PCR) and analyzed for the presence of VK247 or VK210 DNA by oligoprobe hybridization. Sera eluted from a companion filter paper sample were screened for antibodies reactive with the predominant and variant repeat peptides by enzyme-linked immunosorbent assays (ELISA) and with sporozoites by the immunofluorescent antibody (IFA) test. All 24 patients were positive by PCR and oligoprobe hybridization for either VK210 (16 of 24), VK247 (3 of 24), or both (5 of 24). Mixed infections were common (5 of 7) in Peru, but were not observed in the Mexican isolates (0 of 17). All three VK247 infections from Mexico occurred in residents of the foothills above Tapachula (P = 0.02). Of patients with smear-positive P. vivax infection, 42% (10 of 24) had detectable antibodies eluted from dried blood dots that were reactive with the CS protein by IFA or ELISA. These findings establish the widespread distribution of the P. vivax variant CS protein in the New World and indicate that dried blood filter paper samples represent a valuable source of material for the serologic and molecular analysis of plasmodial infections.

Animals↗
PubMed2005

[Molecular and genetic characterization of rifampicin- and/or izoniazid-resistent strains of M. tuberculosis, isolated in Novosibirsk and Tomsk regions].

The purpose of the work was to study rifampicin- and izoniazid-resistent strains of M. tuberculosis, circulating in Western Siberia, by VNTR and IS6110 typing. The authors also studied genetic causes of resistance to these antibiotics and undertook a search of new VNTR loci, displaying polymorphism in genomes of closely related clonally-disseminated variants of Mycobacterium tuberculosis in W-Beijing family model analysis.

Antitubercular Agents↗
PubMed1998

Genetic characterization of accessory genes from human immunodeficiency virus type 1 group O strains.

Human immunodeficiency virus type 1 (HIV-1) group O strains have been described as highly divergent, compared with the vast majority of the viruses involved in the worldwide AIDS pandemic, classified in group M. To gain new insights into the diversity and genetic characteristics of group O, we have sequenced the accessory gene region (from vif to vpu) of 14 isolates. Analyses of the deduced amino acid sequences for Vif, Vpr, the first exon of Tat, and Vpu indicate that most of the functional domains of these proteins, as described for group M viruses, are highly conserved and retained among all the group O strains we have characterized. The only difference concerns the Vif phosphorylation sites, which are absent in all of the group O isolates we have sequenced; in contrast, they are well conserved in nearly all of the group M isolates, in which they play critical roles in the regulation of viral replication and infectivity. As already observed for group M isolates, the Vpu protein is also highly diverse among group O strains. Phylogenetic analyses of these sequences indicate that HIV-1 group O can be separated into four different clusters, containing most of the strains we have characterized (except one, which clusters outside of the analyzed viruses). Taking into account the criteria used for clades in group M, we were not able to define group O clades definitively.

Amino Acid Sequence↗
PubMed1981

Human succinate dehydrogenase: biochemical and genetic characterization.

A simple procedure for the preparation of soluble human succinate dehydrogenase is described. These preparations have proved suitable for analysis by zone electrophoresis, using a specific stain to detect activity after separation. In a survey of succinate dehydrogenase from various tissues and different individuals, no evidence for genetic heterogeneity due to the expression of either multiple loci or alternative alleles at the succinate dehydrogenase locus was found. However, epigenetic heterogeneity in both molecular size and charge was seen and various explanations for the occurrence of the isoenzymes are explored. Estimates of molecular size (93,300 +/- 9100) suggest that the smallest active unit of succinate dehydrogenase accounts for the major part of the solubilized activity. Kinetic studies have shown that the apparent Km values for succinate (0.9 mM) and PMS (0.4 mM) are comparable to those previously described for the beef heart enzyme, and these parameters were not significantly altered when the enzyme was removed from the membrane milieu. However a marked non-succinate-dependent activation of the membrane-associated enzyme at 38 C is apparently lost on solubilization, and this observation may have some bearing on earlier reports of an apparent decrease in Vmax on solubilization of succinate dehydrogenase.

Adult↗