Characterization of hypervariable microsatellite loci in the threespine stickleback Gasterosteus aculeatus.
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Animals↗
Biomedical subjects
U Kuhnlein
Publications and source records attributed to U Kuhnlein.
Endogenous viral gene distribution in populations of meat-type chickens.
The present study was designed to document the complexity of endogenous viral (ev) genes and seek evidence for their association with production traits in selected and control strains of meat-type chickens. Three populations were studied, each consisting of a control strain and one to three strains selected for various production traits. The ev genes were revealed by digesting genomic DNA with restriction enzymes and detecting DNA fragments on Southern blots using radioactive probes from nucleotide sequences of the avian leukosis virus genome. A total of 31 polymorphic ev loci were identified in these populations from a SacI digest, with an average of 7.3 ev genes per bird. There were no significant differences in ev genes per bird between strains within populations or between selected and control strains overall. Thirty of 62 comparisons in the three populations indicated ev gene frequency differences (P less than .05). Within populations, 13 of 93 comparisons of ev gene frequencies between control and selected strains and 8 of 62 between three selected strains of a sire population showed such differences (P less than .05). Selection for body weight and feed efficiency had been observed to reduce gene frequencies of the slow-feathering gene, which usually contains the ev21 locus; however, these effects were not detected (.05 less than P less than .06) between strains of the dam population in the current study. Such differences suggested possible associations between ev genes and production traits in meat-type chickens.
Animals↗
Identification of markers associated with quantitative trait loci in chickens by DNA fingerprinting.
Three approaches for identifying VNTR alleles associated with quantitative traits in chickens are described. One approach is based on the comparison of well-defined selected and non-selected control strains. The second approach is based on analyzing chickens within a breeding population ranked according to specific traits and the third approach involves segregation analysis. In this latter approach a large number of offspring of a single male segregating for a quantitative trait are produced and tested for trait association of the male DNA fingerprinting bands. In all cases pooled DNA samples of birds, rather than individual samples, are analyzed and band intensity is assumed to reflect the relative frequency of an allele. Examples from the literature and from our laboratory indicate that these methods permit the identification of DNA fingerprinting bands associated with quantitative traits. After developing locus-specific probes for these bands it should ultimately be possible to detect and map quantitative trait loci.
Animals↗
Endogenous viral genes: association with reduced egg production rate and egg size in White Leghorns.
Endogenous viral (ev) genes are DNA sequences residing permanently in the genome of most chickens that have a high degree of homology to avian leukosis viruses (ALV). Association of ev genes with production trait differences was studied in White Leghorns free of exogenous ALV. The Cornell Strain K and S chickens used in Experiment 1 had multiple ev genes. In each of four lines of chickens in Experiment 2, there was a 1:1 segregation of full-sibs free of ev genes and those carrying one ev gene: ev-12 that produces the complete endogenous virus, ev-3 or ev-6 that express certain viral antigens, or ev-1, a silent gene. In Experiment 1, the presence of genes ev-10 or ev-19, known to produce the complete virus, was associated with a 9% reduction in the annual egg production rate (P less than .05) in Strain S. Similarly, the presence of the virus-producing ev-12 in Experiment 2 was associated with an 8% reduction of annual egg production rate (P less than .05), a 2.2-g reduction in egg weight (P less than .01), and a .003 reduction in egg specific gravity (P less than .01). No significant effects of ev genes on age at first egg, Haugh unit score, percentage of eggs with blood spots, and body weight of hens were observed. It was concluded that ev genes producing complete endogenous virus are associated with production trait differences similar to those associated with subclinical infections with exogenous ALV.
Animals↗
Nucleotide sequence of turkey prolactin.
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Amino Acid Sequence↗
Increased uracil-DNA glycosylase, AP-DNA binding protein and deoxyribonuclease activities in tumor and SV40-transformed cell lines of human origin.
The activities of three human DNA metabolizing enzymes--uracil-DNA glycosylase, apurinic/apyrimidinic(AP)-DNA binding protein (an AP-DNA endonuclease) and the major cellular deoxyribonuclease (presumably DNase III and/or DNase IV)--were measured in logarithmic growing (diploid non-established) fibroblast strains, tumor-derived cell lines and SV40-transformed cell lines. The levels of activity of uracil-DNA glycosylase and DNase were increased, on average, 5- to 6-fold in tumor cell lines and 10-fold in SV40-transformed cell lines compared to those observed in normal fibroblast strains. AP-DNA binding activity was only 2- to 3-fold higher in both tumor-derived and SV40-transformed cell lines. Measurements in serum-deprived (and hence growth-retarded) SV40-transformed cells indicated that the observed increase in enzyme activity was only partially due to a higher proportion of S-phase cells in the rapidly growing transformed lines. Cell extract mixing experiments indicated that the relatively low levels of activity of the three enzymes in normal fibroblasts could not be ascribed to the presence of an inhibitory factor(s) in the crude extract.
Cell Line↗
Assessment of inbreeding by DNA fingerprinting: development of a calibration curve using defined strains of chickens.
By analyzing DNA fingerprints of chickens from seven well-defined genetic groups, a calibration curve was established relating the degree of inbreeding with the average band frequency, allelic frequency and band sharing. The probe used was bacteriophage M13 DNA and digestion of the genomic DNA was carried out with the MspI restriction enzyme. The analysis also provided an estimate of the average allelic frequency at a hypervariable locus and the average mutation frequency per locus and generation. The values of 0.24 and 1.7 X 10(-3), respectively, are similar to the estimates for humans using other probes and hybridization protocols. It is suggested that the calibration curve established can be used for determining inbreeding not only in chickens, but also in other species.
Alleles↗
Influence of selection for egg production and Marek's disease resistance on the incidence of endogenous viral genes in White Leghorns.
The influence of selection on the frequencies of endogenous viral (ev) genes related to the avian leukosis virus was studied in two genetically distinct sets of White Leghorn strains. Each set consisted of four strains: an unselected control strain, two strains selected for egg production traits, and a strain selected for Marek's disease (MD) resistance as well as egg production traits. Eight different ev genes were observed in Set I and seven in Set II, four being common to both sets. Selection for egg production traits resulted in significant changes of the frequency of four ev genes in both sets. In Set I, increased frequencies were observed for ev-4, ev-7, and ev-8; a decreased frequency for ev-9 was observed. The ev-9 gene expresses the viral envelope protein, whereas the others are transcriptionally silent, with the possible exception of ev-7. In Set II, increased frequencies were observed for the transcriptionally silent ev-8 and for ev-15, a gene which consists of a solitary long terminal repeat. Decreased frequencies were observed for ev-18, which codes for infectious endogenous virus, and for a second ev gene of unknown phenotype. In the resistance-selected strains the frequencies of the ev genes were intermediate between those of the control strains and the strains selected for egg production traits with the exception of ev-6, which expresses the viral envelope protein, and ev-3, which expresses internal viral proteins as well as the envelope protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Comparison of apurinic DNA-binding protein from an ataxia telangiectasia and a HeLa cell line. Evidence for an altered processing of apurinic/apyrimidinic endonuclease.
The major apurinic (AP) DNA-binding protein was purified from a HeLa cell line and from the SV40-transformed cell line AT5BIVA derived from a patient with the repair deficiency syndrome ataxia telangiectasia (AT). This protein appears to be identical with the major cellular apurinic/apyrimidinic endonuclease. The two endonucleases differ in their molecular weight (HeLa, 37,600; AT, 38,900) and their dissociation equilibrium constant for AP sites (HeLa, 7.8 X 10(-11) M; AT, 28.3 X 10(-11) M). These variances might be the consequence of a different post-translational modification. Evidence for this interpretation stems from the observation that the AP DNA binding activity of AP endonuclease, as measured in a glass-fiber filter binding assay, is inactivated upon incubation with snake venom phosphodiesterase and that the AP endonuclease from AT cells in 5-10-fold more sensitive than the HeLa enzyme. For both enzymes, the diesterase treatment leads to the formation of a protein of Mr 35,500 which might be the unmodified precursor of AP endonuclease. The loss of AP DNA binding does not reduce but rather increases the catalytic activity of AP endonuclease when measured at excess substrate concentration.
Ataxia Telangiectasia↗
Cell lines from xeroderma pigmentosum complementation group A lack a single-stranded-DNA-binding activity.
Human fibroblasts and HeLa cells contain two major DNA-binding activities for superhelical DNA, which can be separated by phosphocellulose chromatography. The DNA-binding activity which elutes first from the column coelutes with and is probably identical to a single-stranded-DNA-binding activity. The second activity has been characterized previously. It binds preferentially to super-helical DNA containing DNA damage, but does not bind to single-stranded DNA. Five cell lines derived from patients with the repair-deficiency syndrome xeroderma pigmentosum (XP) were analyzed for the presence of these binding activities. Four of the cell lines were from the A-complementation group and one was from the D-complementation group of XP. The binding activity with preference for damaged DNA was present in all cell lines. The single-stranded-DNA-binding activity was present in the XP-D cell line but was absent or reduced in all of the four XP-A cell lines tested.
Adult↗
The effect of short-term dietary modification on human fecal mutagenic activity.
To assess the effect of short-term modification of diet on human fecal mutagenic activity, 6 subjects consumed 2 dietary regimes hypothesized to affect risk of colorectal cancer. After a 7-day baseline period, a 'low-risk' non-meat diet was consumed for 2 weeks followed by 2 weeks on a 'higher risk' diet which emphasized beef and refined grains. Fecal samples were collected at the end of each diet period and assayed for direct-acting mutagens with the fluctuation test for weak mutagens using Salmonella typhimurium TA100 and TA98 as tester strains. Fecal mutagenic activity on TA100 was increased for all subjects during the 'higher risk' period compared to the 'low risk' period. The average mutagenicity on TA98 was also increased, but the trend was not consistent for all subjects. The baseline diet and non-meat diet resulted in approximately equal mean fecal mutagenicity levels. These findings indicate that a diet high in meat and refined grain, as characterized here, increases fecal mutagenic activity within a 2-week period.
Adult↗
Effects of purified cellulose and pectin fiber diets on mutagenicity of feces and luminal contents of stomach, small and large bowel in rats.
The mutagenicity in extracts of fecal pellets and luminal contents obtained from specific sites along the gastrointestinal tract was examined in male Wistar rats fed a chemically-defined fiber-free diet or nutritionally and calorically equivalent diets containing different amounts of either cellulose or pectin (4.5 or 9.0%) or a mixture of cellulose and pectin (4.5% each). Mutagenicity was measured with the fluctuation test for weak mutagens using Salmonella typhimurium strains TA98 and TA1535 as test organisms. Dietary fiber reduced the mutagenicity of fecal pellets from most of the animals. However, given the number of animals studied, the reduction was only significant for the 9.0% pectin diet (TA98), the 9.0% cellulose diet (TA1535) and the mixed fiber diet (TA1535). Analysis of the contents of stomach, distal small bowel, cecum and colon after a period of 8 weeks on a fiber-free diet revealed that the distal small bowel content had a mutagen concentration that was several times that at any other site. The mutagenicity of the small bowel content of animals on diets containing either 9.0% cellulose or 9.0% pectin was significantly lower than that of animals on a fiber-free diet, as assayed by either of the test strains of S. typhimurium. A similar reduction was detected in animals on the 4.5% cellulose or 4.5% pectin diet by test strain TA1535, but a diet containing a mixture of the same fiber polymers did not reduce the mutagenicity of the small bowel content. The mutagenicity of the contents of the stomach, cecum and colon was not significantly influenced by dietary fiber.
Animals↗
DNA-binding protein from HeLa cells that binds preferentially to supercoiled DNA damaged by ultraviolet light or N-acetoxy-N-acetyl-2-aminofluorene.
A DNA-binding protein was partially purified from extracts of HeLa cells by high-speed centrifugation and chromatography on DEAE-cellulose, phosphocellulose and ultraviolet light-irradiated DNA-cellulose columns. It eluted from the phosphocellulose column with 0.375 M potassium phosphate and from the ultraviolet light-irradiated DNA-cellulose column between 0.5 M and 1 M NaCl. The protein binds preferentially to supercoiled PM2 DNA treated with ultraviolet light or N-acetoxy-N-acetyl-2-aminofluorene, as compared to native supercoiled PM2 DNA. The binding is non-cooperative. Nicked or linear forms of PM2 DNA (damaged or untreated) are not efficient substrates, indicating a requirement of DNA supercoiling for DNA binding. The sedimentation coefficient of the protein estimated by glycerol gradient centrifugation is 2.0-2.5 S, corresponding to a molecular weight of about 20000-25000 if the protein is spherical. The binding to DNA irradiated with ultraviolet light or treated with acetoxyacetylaminofluorene is optimal at around 100-200 mM NaCl and is relatively independent of temperature and pH. MgCl2 and MnCl2 at concentrations between 1 and 5 mM do not markedly affect the binding, but it is inhibited by sucrose, ATP and caffeine. The biological significance of the DNA-binding protein remains to be determined. It does not possess significant glycosylase, endonuclease or exonuclease activities. The dissociation equilibrium constant for the binding reaction of the protein to the ultraviolet light or acetoxyacetylaminofluorene-induced binding sites on DNA is estimated to be 4.10(-11) M. There are at least 1.10(5) DNA-binding protein molecules/HeLa cell.
2-Acetylaminofluorene↗
Mutagens in feces from vegetarians and non-vegetarians.
Mutagens in water extracts from feces of persons in 3 different diet groups were measured with the fluctuation test for weak mutagens using Salmonella typhimurium TA100 and TA98 as tester strains. The 3 diet groups were ovo-lacto vegetarians (N = 6), strict vegetarians (N = 11) and non-vegetarians (N = 12). All subjects were from the urban area of Vancouver, British Columbia, Canada. On TA100 ovo-lacto vegetarians and strict vegetarians had significantly lower levels of fecal mutagens than non-vegetarians (P less than or equal to 0.025 and P less than 0.010, resp.). The same pattern, although less significant, was obtained with TA98. Correlation studies between mutagenicity on TA100 and TA98 and between the pH of the fecal homogenate and mutagenicity indicate the presence of 2 or more major fecal mutagens.
Diet, Vegetarian↗
Cooperative structural transition of PM2 DNA at high ionic strength and its dependence on DNA damages.
Pohl and Jovin have shown that poly(dG--dC) . poly (dG--dC) in solution undergoes a cooperative transition at 2.5 M NaCl at neutral pH. The two forms of the polymer are structurally different as indicated by a change in ethidium bromide intercalation and in circular dichroism (CD). Wang et al. and Davies and Zimmerman have suggested that the low salt form of the polymer might correspond to the B form of DNA (right-handed helix) and the high salt form to the Z form of DNA (left-handed helix). We describe here a salt-induced transition of supercoiled PM2 DNA from a form which passes through nitrocellulose filters to a form which is retained by these filters. The transition occurs between 2.5 and 3.5 M NaCl. The dependence of the apparent equilibrium constant on the salt concentration indicates a cooperative transition. Irradiation of DNA with UV light or alkylation with N-acetoxyacetylaminofluorene (AAAF) shifts the transition to lower salt concentrations. The transition can also be observed with linear DNA but requires a much higher salt concentration.
Acetoxyacetylaminofluorene↗
Disulfiram inhibits DNA breakage by hydroxyradical-producing agents.
Disulfiram (Antabuse, tetraethylthiuram) inhibits DNA cleavage by 1,2-dimethylhydrazine, ascorbate and cysteine. DNA cleavage by these agents is thought to be caused by hydroxyradicals which are formed during auto-oxidation in the presence of transition metals. H2O2 is an intermediate of these auto-oxidation reactions. The inhibitory action of disulfiram is at the level of hydroxyradical formation from H2O2 or at a later step of the reaction, since DNA cleavage by Fenton's reagent (H2O2/FeCl2) is also inhibited. DNA methylation by N-methyl-N'-nitro-N-nitrosoguanidine or methylazoxymethanolacetate is not affected by disulfiram.
Ascorbic Acid↗
Chromosome breaking activity of human feces and its enhancement by transition metals.
Chloroform-methanol extracts from human fecal samples were found to contain compounds which induce chromosome aberrations in Chinese hamster ovary cells. The induction of chromosome aberrations is stimulated by Cu2+ or Mn2+ and inhibited by Fe2+ or Fe3+. Addition of catalase to the fecal extract or the mixture of fecal extract and Mn2+ reduced the frequency of chromosome aberrations. These properties are indicative of hydroxyradical producing agents.
Animals↗
Characterization of DNA damages by filtration through nitrocellulose filters: a simple probe for DNA-modifying agents.
A simple technique for the detection of DNA-modifying agents is described. The double-stranded covalently closed circular DNA of phage PM2 is exposed to the modifying agent and then analysed for DNA damages by assays involving only incubation steps and filtration through nitrocellulose filters. The technique described allows the measurement of DNA modifications which lead to local denaturation of the DNA double helix, interstrand cross-links, single- and double-strand breaks, damages which render the phosphodiester bonds of the DNA sensitive to hydrolysis and damages which labilise the glycosylic bond between base and sugar moiety.
DNA, Circular↗