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E M Prager

Publications and source records attributed to E M Prager.

At least 19 recordsLinked to original sources
PubMed2000

Monitoring Piscirickettsia salmonis by denaturant gel electrophoresis and competitive PCR.

Reported strains of Piscirickettsia salmonis, a pathogen of salmonid fishes, were analyzed by amplifying part of the internal transcribed spacer (ITS) of the ribosomal RNA (rRNA) operon followed by denaturing gradient gel electrophoresis (DGGE) of the amplicons. All amplified fragments differing in sequence were distinguished by migration during DGGE. A simpler format, constant denaturant gel electrophoresis (CDGE), allowed the same diagnostic distinctions among strains. Sampling during 1997 and 1998 of salmonids from 5 different sites on and near Chiloé Island in southern Chile displaying piscirickettsiosis revealed only P. salmonis resembling LF-89, the type strain first isolated in 1989. These observations are encouraging for control strategies, which might otherwise be compromised by unpredictable shifts of P. salmonis types in salmon farms. A competitive PCR assay offered insight about the power of PCR for quantification and about specific tissue invasiveness by this intracellular pathogen. This approach revealed that the PCR could amplify approximately 1 to 10 P. salmonis genome equivalents against a background of > 99.9% salmonid DNA. It also raised the possibility that the salmonid brain is an important site for P. salmonis survival, with its bacterial load in 1 individual having been about 100 times the loads observed in liver and kidney. Pathogen detection by competitive PCR in a surface seawater sample from a netpen in use indicated a density of about 3000 to 4000 P. salmonis cells (or their DNA remnants) 1(-1). Such quantitative estimates should aid decisions about disease prevention and management as, for example, choice of netpen sites following fallow periods and certification of ova, which are known conduits of infection.

Animals↗
PubMed1998

Genetic variation and phylogeography of central Asian and other house mice, including a major new mitochondrial lineage in Yemen.

The mitochondrial DNA (mtDNA) control region and flanking tRNAs were sequenced from 76 mice collected at 60 localities extending from Egypt through Turkey, Yemen, Iran, Afghanistan, Pakistan, and Nepal to eastern Asia. Segments of the Y chromosome and of a processed p53 pseudogene (Psip53) were amplified from many of these mice and from others collected elsewhere in Eurasia and North Africa. The 251 mtDNA types, including 54 new ones reported here, now identified from commensal house mice (Mus musculus group) by sequencing this segment can be organized into four major lineages-domesticus, musculus, castaneus, and a new lineage found in Yemen. Evolutionary tree analysis suggested the domesticus mtDNAs as the sister group to the other three commensal mtDNA lineages and the Yemeni mtDNAs as the next oldest lineage. Using this tree and the phylogeographic approach, we derived a new model for the origin and radiation of commensal house mice whose main features are an origin in west-central Asia (within the present-day range of M. domesticus) and the sequential spreading of mice first to the southern Arabian Peninsula, thence eastward and northward into south-central Asia, and later from south-central Asia to north-central Asia (and thence into most of northern Eurasia) and to southeastern Asia. Y chromosomes with and without an 18-bp deletion in the Zfy-2 gene were detected among mice from Iran and Afghanistan, while only undeleted Ys were found in Turkey, Yemen, Pakistan, and Nepal. Polymorphism for the presence of a Psip53 was observed in Georgia, Iran, Turkmenistan, Afghanistan, and Pakistan. Sequencing of a 128-bp Psip53 segment from 79 commensal mice revealed 12 variable sites and implicated >/=14 alleles. The allele that appeared to be phylogenetically ancestral was widespread, and the greatest diversity was observed in Turkey, Afghanistan, Pakistan, and Nepal. Two mice provided evidence for a second Psip53 locus in some commensal populations.

Animals↗
PubMed1996

Polyclonal antisera elicited bý lysozymes: insights into antigenic structure and evolution.

Polyclonal antisera elicited by evolutionary variants of bird lysozymes c played a major role in the development of the multideterminant-regulatory model to describe the antigenic structure of globular proteins and in the demonstration that there is a strong correlation between immunological differences and amino acid sequence differences. This chapter reviews the evidence and calculations used to show, for lysozyme c and several other proteins, the essentially the entire surface of globular proteins is antigenic, that nearly all evolutionary substitutions affect immunological cross-reactivity, and that there is empirical and theoretical support for the use of immunological distances to infer genealogical relationships and establish approximate evolutionary time scales. In addition this chapter discusses several examples of the use of polyclonal antisera to lysozymes c and g to gain insights into molecular and organismal evolution and the regulation of gene expression.

Animals↗
PubMed1996

Adaptive evolution of lysozyme: changes in amino acid sequence, regulation of expression and gene number.

Adaptive evolution of lysozyme has involved remodelling of amino acid sequences and changes in patterns of gene expression and in gene number. Following an outline of the phenomena likely to be indicative of adaptive evolution and how one can assess them, this chapter focuses on four cases in which lysozyme c has been recruited as a digestive enzyme in the stomachs of creatures needing to retrieve nutrients from microorganisms in fermented food. For each case-ruminant artiodactyls, leaf-eating monkeys, a leaf-eating bird, and fruit flies-the factors likely to be of primary importance in lysozyme's adaptation are examined. Additional examples of apparent adaptation for digestion or antimicrobial defense in animals as diverse as mice, moths, and molluscs are summarized. This chapter considers also the case of three internally clustered residues which among galliform bird lysozymes c occur either as Thr 40, Ile 55, and Ser 91 (TIS) or as Ser 40, Val 55, and Thr 91 (SVT). Reconstruction and testing of six possible intermediate proteins and development of the concept of a neutral corridor of protein traits are described.

Adaptation, Physiological↗
PubMed1996

Animal lysozymes c and g: an overview.

Amino acid sequences for 88 distinct lysozymes c, obtained from members of four vertebrate classes and two orders of insects, are summarized. A model for the relationships and origins of major lineages within the lysozyme c superfamily-which consists of conventional lysozymes c, calcium-binding lysozymes c, and alpha-lactalbumin-is presented and supported by evolutionary analyses. Pioneering events in the discovery and sequencing of lysozymes c are traced, and salient contributions to knowledge made by sequences from various kinds of animals highlighted. A summary of the four known amino acid sequences of bird lysozymes g and an outline of the investigations on this very different kind of vertebrate lysozyme are provided. Areas of future research aimed at further elucidating early events in the evolutionary history of the lysozyme c superfamily and at understanding differences in patterns of lysozyme gene expression are outlined.

Amino Acid Sequence↗
PubMed1996

Mitochondrial DNA sequence variation in the eastern house mouse, Mus musculus: comparison with other house mice and report of a 75-bp tandem repeat.

The control region and flanking tRNAs were sequenced from 139 Mus musculus mitochondrial DNAs (mtDNAs) from mice collected at 44 localities extending from Germany to Japan. Among the 36 types of M. musculus mtDNA resolved, five have an added 75-bp direct repeat; the two copies within an individual differ by two to four base substitutions. Among 90 M. domesticus mtDNAs sequenced, 12 new types were found; 96 M. domesticus types have now been identified by sequencing this segment. Representative mtDNAs from M. castaneus, M. macedonicus, M. spicilegus and M. spretus were also sequenced. A parsimony tree for the M. musculus mtDNAs is about half as deep as the tree for the M. domesticus mtDNAs, which is consistent with the idea that M. musculus is genetically less diverse and younger than M. domesticus. The patterns of variation as a function of position are similar but not identical in M. musculus and M. domesticus mtDNAs. M. castaneus and M. musculus mtDNAs are allied, at a tree depth about three times as great as the start of intra-M. musculus divergence. The coalescence of the M. musculus and M. castaneus mtDNAs is about half as deep as their coalescence with the M. domesticus mtDNA lineages. The mtDNAs of the aboriginal M. macedonicus and M. spicilegus are each other's closest relatives, at a tree depth greater than the deepest intracommensal node. The mtDNA results support the view that the aboriginal M. spretus is the sister group of the other five species.

Animals↗
PubMed1995

Population subdivision and gene flow in Danish house mice.

Genetic subdivision in local populations of the European house mice, Mus musculus domesticus and M. m. musculus, was analysed to study patterns of gene flow. The data consisted of frequencies of microsatellite alleles in 16 samples (250 individuals) from a total of 11 sites in Jutland, which included successive samples from three sites. Sequences of the control region of mitochondrial DNA in three successive samples from one site were also analysed. Microsatellite genotype frequencies within samples were close to Hardy-Weinberg expectations. Levels of microsatellite differentiation among samples (theta = 0.05-0.21) corresponded to limited gene flow at migration-drift equilibrium (Nm = 1-5). Weak isolation by distance for microsatellites in M.m. musculus suggested that gene flow tends to occur among neighbouring sites. Estimates of effective population size over a few generations were much lower than those corresponding to the long periods needed for arrival at mutation-drift equilibrium. This suggested that subpopulations had been influenced by gene flow since formation, or had originated recently from genetically diverse founders.

Analysis of Variance↗
PubMed1993

The sequence-immunology correlation revisited: data for cetacean myoglobins and mammalian lysozymes.

Quantitative microcomplement fixation tests employing rabbit antisera were done to compare immunologically 13 cetacean myoglobins and 15 mammalian lysozymes c of known amino acid sequence. In both cases there was a strong correlation between immunological distance (y) and percent sequence difference (x), as had been found for several other globular proteins. For myoglobin the relationship could be described by y = 10.5x and for lysozyme by y = 8.5x. The coefficients in both of these equations are appreciably higher than the values of 5.1-6.9 reported for three other vertebrate globular proteins (bird lysozyme c, mammalian ribonuclease, and mammalian serum albumin), and they imply that rabbit antisera to mammalian myoglobins and lysozymes are more sensitive to evolutionary substitutions. A strong inverse correlation (r = -0.95) was found when the slope of the line relating y to x for these five data sets was plotted against the percent sequence difference between the rabbit's own protein and the proteins immunized with. Specifically, the cetacean myoglobins on average differ in amino acid sequence from rabbit myoglobin by less than 13% and exhibit the steepest slope (10.5), while bird lysozyme sequences differ by nearly 40% from rabbit lysozyme and exhibit the shallowest slope (5.1).

Amino Acid Sequence↗
PubMed1993

Mitochondrial DNA affinity of several Jewish communities.

The mitochondrial DNA (mtDNA) of 332 individuals from Israel, including 270 Jews (originating from 7 communities) and 62 Arabs, was analyzed. Each mtDNA haplotype was determined by the fragment patterns of restriction enzymes HpaI, BamHI, HaeII, MspI (HpaII), and AvaII. The variability of the total sample and of each community was high. Of 40 different haplotypes, 20 were found more than once. Most haplotypes are typical of Caucasians, but African types were found among Ethiopian Jews and to a lesser extent among Arabs. The communities differed in their haplotypes: Chi-square tests among six communities showed significant differences for most pairwise comparisons and nonsignificant differences involving mainly the Moroccan Jews. In a genetic distance analysis only the Ethiopian Jews appeared to be distinguished from the other communities. According to a GST analysis, approximately 30% of the variation among the mtDNA restriction maps is attributable to differences between communities.

Chi-Square Distribution↗
PubMed1992

Evolutionary genetics of ruminant lysozymes.

Comparative studies of mammalian lysozymes and their genes have contributed to knowledge of how new functions arise during evolution. The recruitment of lysozymes for functioning in the stomach fluid of ruminants has occurred in response to selection pressures that are partly known and on a time-scale that is known. A semiquantitative analysis of adaptive evolution is thus made possible by the ruminant lysozyme system. Large-scale production of lysozyme by the stomach lining entailed gene duplication as well as a change in gene expression. Remoulding of the lysozyme for working and lasting in the stomach fluid involved accelerated amino acid replacements, which may have been facilitated by intergenic recombination. The possibility that multigene families can accelerate adaptive evolution, by virtue of their capacity for bringing together functionally coupled substitutions, receives emphasis in this review.

Adaptation, Physiological↗
PubMed1991

cDNA and amino acid sequences of rainbow trout (Oncorhynchus mykiss) lysozymes and their implications for the evolution of lysozyme and lactalbumin.

The complete 129-amino-acid sequences of two rainbow trout lysozymes (I and II) isolated from kidney were established using protein chemistry microtechniques. The two sequences differ only at position 86, I having aspartic acid and II having alanine. A cDNA clone coding for rainbow trout lysozyme was isolated from a cDNA library made from liver mRNA. Sequencing of the cloned cDNA insert, which was 1 kb in length, revealed a 432-bp open reading frame encoding an amino-terminal peptide of 15 amino acids and a mature enzyme of 129 amino acids identical in sequence to II. Forms I and II from kidney and liver were also analyzed using enzymatic amplification via PCR and direct sequencing; both organs contain mRNA encoding the two lysozymes. Evolutionary trees relating DNA sequences coding for lysozymes c and alpha-lactalbumins provide evidence that the gene duplication giving rise to conventional vertebrate lysozymes c and to lactalbumin preceded the divergence of fishes and tetrapods about 400 Myr ago. Evolutionary analysis also suggests that amino acid replacements may have accumulated more slowly on the lineage leading to fish lysozyme than on those leading to mammal and bird lysozymes.

Amino Acid Sequence↗
PubMed1991

mtDNA polymorphism in two communities of Jews.

Twenty-one types of mtDNA were found in a survey of 39 Israeli Jews, of whom 18 were Sephardic and 21 Ashkenazic. The survey was made with six restriction enzymes that together recognize an average of 600 bp/genome. The differences among the types appear to be due to base substitution at 19 cleavage sites, one deletion, and one conformational mutation. The numbers of differences imply that these modern Jews stem from a minimum of 21 maternal lineages that were already distinct from one another 4,000-5,000 years ago. In three of the four cases where a type was found in more than one person, it occurred in both Ashkenazic and Sephardic populations. The diversity of types in the combined sample of two Jewish populations is lower than both that in a sample from various parts of Africa and that in a sample from various parts of East Asia. Nevertheless, it is as high as that in a sample from diverse parts of New Guinea, an area much larger than that in Israel to which the Jewish population traces back.

Base Composition↗
PubMed1990

Amino acid sequences of stomach and nonstomach lysozymes of ruminants.

Complete amino acid sequences are presented for lysozymes c from camel and goat stomachs and compared to sequences of other lysozymes c. Tree analysis suggests that the rate of amino acid replacement went up as soon as lysozyme was recruited for the stomach function in early ruminants. The two lysozymes from goat stomach are the products of a gene duplication that probably took place before the divergence of cow, goat, and deer about 25 million years ago. Partial sequences of three lysozymes from goat tears indicated that (a) the goat tear family of lysozymes may have diverged from the stomach lysozyme family by an ancient duplication and (b) later duplications are probably responsible for the multiple forms of tear and milk lysozymes in ruminants.

Amino Acid Sequence↗
PubMed1989

Episodic evolution in the stomach lysozymes of ruminants.

By sequencing lysozymes c from deer and pig stomachs and comparing them to the known amino acid sequences of other lysozymes c, it was possible to examine the rate of sequence change during and after the period in which this enzyme acquired a new function. Evolutionary tree analysis suggests that the rate went up while lysozyme was being recruited to function as a digestive enzyme in the stomach of early ruminants. Later, presumably after lysozyme was well adapted for functioning in the new environment, which contains acid, pepsin, and fermentation products, the rate of amino acid replacement became subnormal.

Adaptation, Physiological↗
PubMed1988

Ancient origin of lactalbumin from lysozyme: analysis of DNA and amino acid sequences.

Parsimony trees relating DNA sequences coding for lysozymes c and alpha-lactalbumins suggest that the gene duplication that allowed lactalbumin to evolve from lysozyme preceded the divergence of mammals and birds. Comparisons of the amino acid sequences of additional lysozymes and lactalbumins are consistent with this view. When all base positions are considered, the probability that the duplication leading to the lactalbumin gene occurred after the start of mammalian evolution is estimated to be 0.05-0.10. Elimination of the phylogenetic noise generated by fast evolution and compositional bias at third positions of codons reduced this probability to 0.002-0.03. Thus the gene duplication may have long preceded the acquisition of lactalbumin function.

Animals↗
PubMed1987

Recruitment of lysozyme as a major enzyme in the mouse gut: duplication, divergence, and regulatory evolution.

Two major types of lysozyme c (M and P) occur in the mouse genus, Mus, and have been purified from an inbred laboratory strain (C58/J) of M. domesticus. They differ in physical, catalytic, and antigenic properties as well as by amino acid replacements at 6 of 49 positions in the amino-terminal sequence. Comparisons with four other mammalian lysozymes c of known sequence suggest that M and P are related by a gene duplication that took place before the divergence of the rat and mouse lineages. M lysozyme is present in most tissues; achieves its highest concentration in the kidney, lung, and spleen; and corresponds to the lysozyme partially sequenced before from another strain of M. domesticus. In M. domesticus and several related species, P lysozyme was detected chiefly in the small intestine, where it is probably produced mainly by Paneth cells. A survey of M and P levels in 22 species of muroid rodents (from Mus and six other genera) of known phylogenetic relationships suggests that a mutation that derepressed the P enzyme arose about 4 million years ago in the ancestor of the housemouse group of species. Additional regulatory shifts affecting M and P levels have taken place along lineages leading to other muroid species. Our survey of 187 individuals of wild house mice and their closest allies reveals a correlation between latitude of origin and level of intestinal lysozyme.

Amino Acid Sequence↗