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R K Selander

Publications and source records attributed to R K Selander.

At least 127 records · Page 7Linked to original sources

Characterization of a monoclonal antibody to human alpha-fetoprotein and its use in affinity chromatography.

A hybrid cell line secreting monoclonal antibodies to human alpha- fetoprotein (AFP) was produced by fusion of a mouse myeloma cell line with spleen cells from a BALB/c mouse immunized with human AFP. The affinity constant of the antibody was about 1.7 X 10(9) l/mol. When clones were grown in vitro, the highest concentration of specific antibody in the culture medium was 25 microgram/ml. A clone was transplanted intraperitoneally into pristine-primed BALB/c recipients. Ascites developed within 3-4 weeks of transplantation, and the maximal antibody concentration in the ascitic fluid was 2.5 mg/ml. The immunoglobulin fraction of ascitic fluid was coupled to cyanogen bromide-activated Sepharose and used for affinity chromatography of AFP. AFP containing less than 1% contaminating proteins was obtained by passing amniotic fluid through the column and eluting the adsorbed AFP with 4 mol/l urea. The monoclonal antibody was used for radioimmunoassay (RIA). The sensitivity obtained was 50 microgram/l, which is adequate for certain clinical applications.

Animals↗

Genetic diversity and temporal variation in the E. coli population of a human host.

Electrophoretic techniques were employed to study variation in chromosomal genes encoding enzymes and in the distribution of cryptic plasmids in the E. coli population of a human host over an 11-month period. Thirteen of the 15 enzymes studied were polymorphic, and mean genetic diversity per locus was 0.39. Among 550 clones isolated from fecal samples, protein electrophoresis revealed 53 distinct electrophoretic types (ETs). Most ETs appeared on only one or a few days and were considered transients, but two (ET-12 and ET-13) were observed many times over extended periods and represented residents. Complete turnover in the transient ETs in the population occurred in periods of from two weeks to a month. ETs appearing in one month showed no particular genetic similarity to those of the previous month. - All but 4 of the 53 ETs carried one or more "cryptic" plasmids with molecular weights ranging from 1 to 80 megadaltons. With few exceptions, the plasmid composition of each ET was unique. In the course of the 11-month sampling period, there were changes in the plasmid profiles of the resident strains ET-12 and ET-13, and also in the profile of a recurrent strain, ET-2, which was isolated on four days. Modification of the plasmid profile of ET-12 involved the sequential addition of relatively high molecular weight bands. For ET-2 and ET-13, the changes in the plasmid profiles were radical, suggesting invasions of new cell types rather than merely the addition and deletion of plasmids. - The results of this study provide three lines of evidence that recombination plays a minor role in the generation of genetic diversity in the E. coli population of a single host. (1) Several pairs of loci were in strong linkage disequilibrium; compared to a randomly generated array of genotypes, the sample of ETs contained an excess of pairs differing at one or two loci and too many pairs with highly distinctive combinations of electromorphs. (2) In most cases where pairs of ETs differed at a single locus and, therefore, could reasonably have been generated by phage- or plasmid-mobilized gene transfer, the plasmid profiles of the pair members were radically different and/or the potentially transmitted alleles were not present in other ETs in the population. (3) Although ET-12 was abundant, being represented by 252 of the 550 clones sampled, the electrophoretic type most similar to ET-12 different from it at six loci, and ET-12 carried two unique alleles. We conclude that most of the genetic diversity observed in this human host is a consequence of successive invasions of E. coli genotypes.

Alleles↗

Genetic diversity and structure in Escherichia coli populations.

A survey of electrophoretic variation in 20 enzymes from 109 clones of escherichia coli from natural populations yielded an estimate of mean genetic diversity approximately twice that reported in an earlier study and four to five times larger than estimates fro most eukaryotic species. Despite this extensive variability, the number of distinctive genotypes apparently is rather limited. Identical clones were obtained from unassociated hosts, and a clone that is electrophoretically indistinguishable from the laboratory strain Escherichia coli K-12 was isolated from a human infant. The results suggest that rates of genetic recombination in natural populations of Escherichia coli are low. These findings have implications for our understanding of the genetic structure of Escherichia coli populations and the factors determining the amount of neutral gene variability in this bacterial species.

Biological Evolution↗

Self-fertilization and monogenic strains in natural populations of terrestrial slugs.

Electrophoretic studies of genetic variation in 14 species of terrestrial slugs of the families Arionidae, Philomycidae, and Limacidae in the eastern United States indicate that self-fertilization, either facultative or obligatory, is the normal breeding system in six of the species. Three of these six species are single monogenic strains; one consists of three monogenic strains; one includes a highly heterozygous form and two monogenic strains; and one has a moderate amount of polymorphism but little heterozygosity and strong linkage disequilibrium. Eight species are outcrossers, being highly polymorphic and panmictic within local populations. Niche breadth, assessed in terms of extent of geographic distribution and variety of habitats occupied and measured on an experimental plot of woodland, is greater in some monogenic strains than in highly heterozygous, outcrossing species. Colonizing success apparently is independent of the amount of genetic variation carried by a species.

Journal Article↗

Estimating total genic diversity in the house mouse.

In a survey of variation in both electrophoretic charge and thermostability at 14 structural loci in 40 strains of Mus musculus, 27 electromorphs (polypeptides differing in electrophoretic charge) and 20 thermomorphs (polypeptides differing in thermostability) were found. Electrophoresis detected 11 new variants within thermomorphs, and the heat denaturation technique detected four new variants within electromorphs. From these data, and making the assumption that both techniques are independent of each other, it is estimated that the actual total number of alleles at the 14 loci is 53, or an average of 3.79 per locus (1.96 per electromorph), and that electrophoresis apparently detects one-third of the variants, thus describing about 50% of the alleles at structural gene loci in the house mouse.

Alleles↗

Systemic lupus erythematosus and DNA antibodies in pleural effusions.

The quantity of antibodies to double-stranded DNA (ds-DNA) in 53 pleural effusions from 48 patients was measured by means of a modified Farr technique. In 10 samples, binding of ds-DNA was greater than 5 mg/l (range 6--14 mg/l), five samples being from patients with systemic lupus erythematosus (SLE), four from patients with lung cancer, and one from a patient with pulmonary tuberculosis. After treatment of pleural effusion samples with DNase, there was a marked increase of ds-DNA binding in the SLE group (n = 5), but none in the lung cancer group (n = 7) or in 4 patients with pleural effusions of various origin. In pleural fluid, demonstration of antibodies to ds-DNA and anti-ds-DNA-ds-DNA complexes, unmasked by DNase, may prove valuable when differentiating clinical conditions with pleural effusions.

Antibodies, Antinuclear↗

The organization of genetic diversity in the parthenogenetic lizard Cnemidophorus tesselatus.

The parthogenetic lizard species Cnemidophorus tesselatus is composed of diploid populations formed by hybridization of the bisexual species C. tigris and C. septemvittatus, and of triploid populations derived from a cross between diploid tesselatus and a third bisexual species, C. sexlineatus. An analysis of allozymic variation in proteins encoded by 21 loci revealed that, primarily because of hybrid origin, individual heterozygosity in tesselatus is much higher (0.560 in diploids and 0.714 in triploids) than in the parental bisexual species (mean, 0.059). All triploid individuals apparently represent a single clone, but 12 diploid clones were identified on the basis of genotypic diversity occurring at six loci. From one to four clones were recorded in each population sampled. Three possible sources of clonal diversity in the diploid parthenogens were identified: mutation at three loci has produced three clones, each confined to a single locality; genotypic diversity at two loci apparently caused by multiple hybridization of the bisexual species accounts for four clones; and the remaining five clones apparently have arisen through recombination at three loci. The relatively limited clonal diversity of tesselatus suggests a recent origin. The evolutionary potential of tesselatus and of parthenogenetic forms in general may be less severely limited than has generally been supposed.

Alleles↗

Molecular basis of chromosome banding. I. The effect of mouse DNA fractions on two fluorescent dyes in vitro.

The effects of mouse satellite, main band and total DNA on the fluorescence intensity of quinacrine and of the bibenzimidazole derivative Hoechst 33258 were tested in solution. No significant differences were noticed between the double-stranded DNAs in spite of the 5% difference in AT-content between satellite and main band DNA. Single-stranded DNAs enhanced the fluorescence intensity of Hoechst 33258 far less than double-stranded DNAs. Having been denaturated and then reassociated the DNA fractions were intermediate in their enhancing effects on the fluorescence intensity of Hoechst 33258, the differences presumably being due to different degrees of reassociation. The effect of denatured and subsequently reassociated satellite DNA on the fluorescence intensity of quinacrine was similar to that of the native DNAs. Main band and total DNA quenched the fluorescence intensity of quinacrine more after denaturation-reassociation than it did when native. In the discussion the results are related to known cytological data.

Animals↗

Molecular basis of chromosome banding. II. The effect of silver and mercury ions on the fluorescence intensity of acranil-DNA complexes.

Silver and mercury ions are known to react with the bases of nucleic acids in solution. At low cation/base ratios Ag+ has an affinity for GC pairs in DNA, whereas Hg++ is preferentially bound to AT-rich nucleic acids. We have used fluorometry to measure the effect of these cations on the fluorescence intensity of preformed complexes of acranil and DNA in solution. The results are: 1) Ag+ enhances the fluorescence intensity presumably by affecting the dye intercalated in the vicinity of GC-pairs. 2) The addition of Hg++ leads to a quenching of the fluorescence intensity of the complex at low ion/base ratios, suggesting an effect on the dye molecules bound to AT pairs. At high GC-content of the nucleic acid, slight enhancement of the fluorescence intensity occurs with Hg++. 3) With both metals there is a correlation between base content of DNA and effect on the intensity of fluorescence indicating base specificity of the dye-polymer interaction.

Acridines↗

Trophic radiation through polymorphism in cichlid fishes.

Several morphologically defined species of cichlid fishes (Cichlasoma) endemic to the Cuatro Cienegas basin of Mexico and differing in tooth structure, body shape, and diet are allelically identical at 27 gene loci. The presence of only one Mendelian population in each of three drainage systems studied and the occurrence of two of the morphotypes in the same broods indicate that the supposed species are morphs. That trophic radiation in the Cuatro Cienegas cichlids has been achieved through ecological polymorphism rather than speciation raises questions regarding the genetic basis for the extensive intralacustrine radiation of cichlids in Africa and elsewhere.

Journal Article↗

Elephant seals: genetic variation and near extinction.

Blood samples from northern elephant seals (Mirounga angustirostris), representing five breeding colonies in California and Mexico, were surveyed electrophoretically for protein variation reflecting underlying genetic differences. No polymorphisms were found among 21 proteins encoded by 24 loci. This uniform homozygosity may be a consequence of fixation of alleles brought about by the decimation of this species by sealers in the last century.

Alleles↗