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M Georges

Publications and source records attributed to M Georges.

At least 73 records · Page 4Linked to original sources

Human variable number of tandem repeat probes as a source of polymorphic markers in experimental animals.

Human VNTR (Variable Number of Tandem Repeat) markers are examined as a source of polymorphism for linkage studies in inbred strains of mice and rats. High frequencies of cross-hybridization are found under fingerprinting conditions that detect many distinct minisatellite loci in these species. Linkage studies suggest that minisatellite markers are widely distributed in the mouse genome, in contrast to humans where they are clustered, particularly in telomeric regions. Human VNTR probes can be used to screen in mouse genomic libraries to isolate mouse specific VNTR sequences. Some of these sequences reveal fingerprint patterns under stringent hybridization conditions.

Animals↗

Characterization of a set of variable number of tandem repeat markers conserved in bovidae.

Screening purpose-built libraries with minisatellite probes, we have isolated 36 bovine variable number of tandem repeat markers (VNTRs) characterized by a mean heterozygosity of 59.3 within the American Holstein breed. Matching probabilities and exclusion powers were estimated by Monte-Carlo simulation, showing that the top 5 to 10 markers could be used as a very efficient DNA-based system for individual identification and paternity diagnosis. The isolated VNTR systems should contribute significantly to the establishment of a bovine primary DNA marker map. Linkage analysis, use of somatic cell hybrids, and in situ hybridization demonstrate that these bovine VNTRs are scattered throughout the bovine genome, without evidence for proterminal confinement as in the human, and that at least some of them are organized as clusters. Moreover, Southern blot analysis and in situ hybridization demonstrate conservation of sequence and map location of minisatellites within Bovidae.

Animals↗

Generation of bovine multisite haplotypes using random cosmid clones.

One hundred ten random cosmids were used to probe Southern blots of DNA from nine unrelated cattle digested with 12 restriction enzymes. Although only one-third of the expected fragments were explored, 85% of the cosmids revealed at least one polymorphism. The mean heterozygosity of the generated haplotypes was estimated at 51.9%. A surprisingly high proportion of polymorphisms (approximately 25%) was attributed to insertion-deletion events, compensating for the lower level of nucleotide diversity observed in cattle (pi approximately 0.0007) compared to that in human. The mutation rate at cytosines in the CpG dinucleotide was estimated approximately 10 times higher than that at other nucleotides. When used in linkage studies, the generated markers should cover approximately 50% of the bovine genome.

Animals↗

The conservation of dinucleotide microsatellites among mammalian genomes allows the use of heterologous PCR primer pairs in closely related species.

The high degree of polymorphism displayed by DNA microsatellites makes them useful as DNA markers in linkage studies. A search of the DNA sequence databases revealed that the locations of dinucleotide microsatellites are often conserved among mammalian species, enabling the prediction of the presence of DNA microsatellites using comparative genetic data. In closely related species such as cattle and sheep, this conservation was close enough to allow PCR primers designed for use in one species to be used to analyze microsatellite length polymorphism in the other. A total of 48 sets of primer pairs, flanking bovine microsatellites and giving polymorphic PCR products in that species, were tested with template DNA from sheep, horses, and humans. Specific products were obtained in 27 cases (56%) with ovine DNA, 20 of which (42%) showed polymorphisms. With equine DNA, 3 (6.2%) gave specific but monomorphic products, while no specific products were obtained using human DNA. The ability to use heterologous PCR primers, coupled with comparative mapping information will facilitate the use of DNA microsatellites in gene mapping studies in closely related species such as cattle and sheep, rat and mouse, or primates.

Animals↗

On the use of DNA fingerprints for linkage studies in cattle.

To find a marker for the bovine "muscular hypertrophy" gene and for the "roan" locus, we have typed six cattle pedigrees totaling 540 animals for nine blood group systems, for 12 biochemical markers, for RFLPs at four loci, and with five probes revealing multilocus DNA fingerprints. Segregation analysis of the fingerprint bands showed that, in cattle, a fingerprint probe will reveal a mean of 7.6 clearly resolvable bands, behaving as simple, highly informative Mendelian entities characterized by a mean mutation rate of +/- 1/4500 gametes. For one of the bands, we observed a "mutation burst" generating germline mosaicism. Because some of the fingerprint bands were allelic or corresponded to clustered minisatellites, a mean of only 5.7 independent loci is explored per probe. Fingerprint bands revealed by different probes also show a clear propensity for close linkage, pointing toward nonrandom distribution of minisatellite sequences or the existence of minisatellite clusters. Although this reduces the power of fingerprints for linkage analysis substantially, we were able to demonstrate genetic linkage between fingerprint bands and at least three of the classical markers, to exclude the roan locus from 4.5 Morgans of the bovine genome with the DNA fingerprints and for an additional 2.5 Morgans with the classical markers, and to identify a solid candidate marker for the bovine muscular hypertrophy gene, yielding a lod score greater than or equal to 2.84 without any obliged recombinant.

Animals↗

A polymorphic satellite sequence maps to the pericentric region of the bovine Y chromosome.

Exploiting a serendipitously observed bovine male-specific signal, generated by the mouse pSP64.2.5EI minisatellite probe, we have cloned a bovine (Bos taurus) Y-specific sequence: btDYZ-1. This sequence is composed of 60 tandem repetitions of a motif consisting of two parts: a 40-bp-long unit, showing a mean divergence of 27% between repeats, separated from the next repeat by a TG-rich stretch varying in length between 12 and 63 bp. The number of copies of this repeated motif has been estimated at 6 X 10(4) per male genome. As a consequence, the corresponding satellite, DYZ-1, might represent approximately 1/20 of the bovine Y chromosome. btDYZ-1 has been mapped by in situ hybridization to the pericentric region of the Y chromosome. It is characterized by a substantial genetic polymorphism and has been shown to be conserved within the Bos and Bison genera of the Bovinae subfamily. This sequence is being used to develop a sexing procedure for bovine preimplantation embryos based on the polymerase chain reaction.

Animals↗

Structure, evolution and chromosomal localization of the human pregnancy-specific beta 1 glycoprotein gene family.

Cloning and sequencing of the cDNA of a new member of the PS beta G gene family is reported. Comparison of the sequence with those of other PS beta G cDNAs reveals a remarkable conservation of their sequence (greater than 90%) and of their general structural organization: an NH2 domain is followed by 93- and 85-residue Ig-like domains termed A and B, respectively. Most PS beta G contain two domains A in tandem followed by a single domain B. In some PS beta G members described here, alternative splicing skips the AI domain in some transcripts, yielding two- and three-domain variants, respectively. Individual PS beta G members have specific short carboxyl domains displaying little sequence conservation. The PS beta G family is closely related to the CEA gene family. A detailed comparison of the sequence of both families is given and used to construct an evolutionary tree, using the method of Li, Wu, and Luo (1985, Mol. Biol. Evol. 2: 150-174). Computation of the number of substitutions of synonymous (Ks) and nonsynonymous (Ka) sites and of the Ks/Ka ratio suggests that the PS beta G gene family appeared concomitantly with the expansion of the placental mammals and belongs to the class of rapidly evolving genes. Very little selective pressure has been exerted on the body of the molecules, especially on domain A. The analysis also suggests that PS beta G genes encoding different carboxyl domains would have been positively selected and fixed during the evolution. The PS beta G gene family was assigned to chromosome 19, which also carries the CEA genes.

Amino Acid Sequence↗

Minisatellite linkage maps in the mouse by cross-hybridization with human probes containing tandem repeats.

Tests of 29 human variable number of tandem repeat probes in inbred mouse lines showed that 80% (23/29) cross-hybridize, and 48% (14/29) produce multiple band, minisatellite polymorphisms (fingerprint patterns). Mini-satellite-type polymorphisms detected by 11 probes were characterized in eight different strains; on average, 240 polymorphic differences were detected between pairs of strains. Reproducible fingerprint patterns permit the study of the segregation of the minisatellite polymorphisms in experimentally designed crosses. As an example, we constructed primary minisatellite genetic linkage maps containing 346 polymorphic bands, distributed in 101 groups of closely linked systems, from genotypes on a recombinant inbred panel (C57BL/6J X DBA/2J); 38 of the groups were assigned by linkage to 15 autosomal chromosomes. The minisatellite genetic maps of C57BL/6J and DBA/2J can be applied in other linkage studies involving these strains.

Animals↗

The measurement of principled morality by the Kohlberg Moral Dilemma Questionnaire.

The four stages preceding the postconventional level in the Kohlberg (1958, 1971, 1976) system of moral development are described as involving moral judgments that conform to external conditions of punishment, reward, social expectation, and conformity to the law. No special level of self-control seems necessary to behave in keeping with these conditions of external reinforcement. In contrast, the two stages of postconventional (principled) mortality involve defiance of majority opinion and defiance of the law--actions that would seem to require greater self-control. This study was concerned with whether postconventional moral reasoning, as measured by the Kohlberg Moral Dilemma Questionnaire (MDQ), can be associated with higher self-control. If so, prediction of principled moral behavior from the MDQ would be based not only on postconventional moral reasoning but bolstered by the necessary level of self-control as well. College students who came the closest to postconventional moral reasoning showed better self-control than college students who were more conventional or preconventional in their moral judgments. These results support the validity of the MDQ for predicting principled moral behavior.

Adolescent↗

Linkage relationships among 20 genetic markers in cattle. Evidence for linkage between two pairs of blood group systems: B-Z and S-F/V respectively.

Five bovine paternal half-sib pedigrees for a total of 527 individuals were typed for six blood group systems: A, B, F/V, L, S, Z; for nine biochemical polymorphisms: ADA, MPI, PGM-3(slow), NP, Gc, Pi2, Tf, Ptf1 and Ptf2; and for restriction fragment length polymorphisms at five autosomal loci: Tg, GH, LDLr, BoLA-DQ and BoLA-DY. Two of the pedigrees were informative for segregation at the 'muscular hypertrophy' locus, and one was informative at the coat colour determining 'roan' locus. Linkage analysis was performed between all markers. Linkage was demonstrated between the S and F/V blood group systems (z = 3.11), adding one locus to the previously identified linkage group VII (LGVII) [Pi-2 and S], the most likely order being Pi2-S-F/V with maximum likelihood recombination rates of 0.208 and 0.211. Also shown to be linked were the blood group systems B and Z (z = 5.7, theta = 0.245). We confirmed the observation previously made by Andersson et al. (1988) of a high recombination rate between class II genes DQ and DY, suggesting either a larger physical distance between those genes than expected from comparative data, or the presence of a 'recombinational hotspot' in the bovine major histocompatibility complex. No linkage was found either with the 'muscular hypertrophy' locus, or with the 'roan' locus. However, these two loci could be excluded from respectively 1.7 and 2.5 Morgans of the bovine genome.

Animals↗

Demonstration of spontaneous XX/XY chimerism by DNA fingerprinting.

The M13 bacteriophage probe, which makes possible the establishment of DNA fingerprints, was used to study a phenotypically normal woman with a 46XY karyotype and her twin brother. Identical fingerprints and positive hybridization with a series of Y-specific probes were obtained on blood samples from the siblings. DNA from a skin biopsy of the woman yielded a clearly different pattern and displayed no Y-specific hybridization, indicating that she is a spontaneous chimera. This study illustrates the use of DNA fingerprinting as a powerful and simple aid to the diagnosis of natural chimerism.

Adult↗

The bovine gene map.

The present status of the bovine gene map as well as some of the methods and strategies important for future efforts in completing the gene map of cattle are reviewed.

Animals↗

Analysis of genetic variation in the Belgian Blue cattle breed using DNA sequence polymorphism at the growth hormone, low density lipoprotein receptor, alpha-subunit of glycoprotein hormones and thyroglobulin loci.

New DNA sequence polymorphisms were identified at four bovine autosomal loci: growth hormone, low density lipoprotein receptor, alpha-subunit of glycoprotein hormones and thyroglobulin. Assuming independent assortment between these polymorphisms, the probabilities to be heterozygous at these four loci are 0.48, 0.36, 0.10 and 0.77 respectively, within the Belgian Blue Cattle breed (BBCB). Nucleotide diversity was estimated, showing that animals from the BBCB are heterozygous for 1/1450 nucleotides, a value significantly lower than the 1/500 value found in man. Moreover, we have estimated that the mutation rate at the cytosines of CG dinucleotides is about 10 times higher than that for other nucleotides.

Alleles↗

Use of DNA bar codes to resolve a canine paternity dispute.

The DNA fingerprinting method was used to resolve a canine paternity dispute. During the same estrus, a Shih Tzu bitch was inseminated by 2 dogs--a Shih Tzu and a Coton de Tulear. Because both breeds are alike phenotypically, it was difficult to decide whether the pups were purebred or of mixed breeding. The DNA bar codes indicated unambiguously that the 2 sires had fathered one pup each, thus documenting superfecundation.

Alleles↗

DNA fingerprinting in domestic animals using four different minisatellite probes.

Four probes known to allow DNA fingerprinting in the human (M13, Jeffreys' core sequence, the human alpha globin hypervariable region [HVR], and a mouse probe related to the Drosophila Per gene) were checked for their ability to reveal "genetic bar codes" in cattle, horses, pigs, dogs, chickens, and a European cyprinid fish, the barbel (Barbus barbus L.). Individual-specific patterns were obtained in cattle using M13, Jeffreys' core sequence, and the alpha globin HVR, in horses, dogs, and pigs using M13, Jeffreys' core sequence, and the Per probe, and in chicken and fish using the four different probes. Although we observed a considerable heterogeneity in the extent of interindividual variation, depending on the particular probe-species combination, the fingerprints are polymorphic enough to be used efficiently in animal identification, paternity testing, and as a source of genetic markers for linkage analysis. These markers should substantially accelerate the mapping of genes affecting economically important traits.

Animals↗