Genetics and Genomics Commons^™

Characterization Of Hsd::Mudx128 Operon Fusion Mutant Of Escherichia Coli K-12, Marjorie Ann T. Reyno

Loma Linda University Electronic Theses, Dissertations & Projects

The study of the regulation of gene expression in complex genes can be facilitated by the use of operon fusions which place a well characterized lacZ gene under the control of a promoter of interest. An hsd::MudX128 operon fusion mutant of E. coli K-12 isolated by Prakash (1986) was observed to have a high β-galactosidase activity (1000 units), ten to twenty times higher than other hsd::MudX mutants. This β-galactosidase activity was suppressed to a very low level (25 units) by the introduction of an F’ plasmid, F'101, which carries the 98 min to 2 min region of the …

Macrophages But Not B Cells From Aged Mice Are Defective In Stimulating Autoreactive T Cells In Vitro, Aruna Seth, Mitzi Nagarkatti, Prakash S. Nagarkatti, Subbarao Bondada, Venkatachalam Udhayakumar

Microbiology, Immunology, and Molecular Genetics Faculty Publications

In the present study the effect of aging on the capacity of Ia⁺ cells to stimulate autoreactive T cells in the syngeneic mixed lymphocyte reaction (SMLR) was investigated. Using young CD4⁺ T cells as responders, it was observed that unseparated whole spleen cells from aged mice had normal stimulatory activity comparable to that of young spleen cells. Interestingly, however, when purified splenic adherant cells (SAC) enriched for macrophages or splenic B cells were used as stimulators, aged SAC but not aged B cells were found to be defective in stimulating autoreactive T cells. This defect in aged SAC …

Phenotypic And Functional Properties Of B Lymphocytes From Aged Mice, Subbarao Bondada, Joan Morris, Richard J. Kryscio

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Phenotypic and functional properties of B lymphocytes from individual young and old mice of different inbred strains were studied. B lymphocyte subpopulation defined by the ratios of the densities of cell surface IgM and IgD were found to be altered with age. However, such alterations in B cell subsets were found only in 30-40% of the old mice. B cell mitogenic responses to anti-μ and anti-Lyb2 antibodies were decreased in a majority of DBA/2 mice. Proliferative responses to LPS and antiμ were reduced only in a minority of CBA/Ca mice but there was a very good correlation in the responsiveness …

Micronuclear Genome Organization In Euplotes Crassus: A Transposonlike Element Is Removed During Macronuclear Development, Scott Everet Baird, Gina M. Fino, Susan L. Tausta, Lawrence A. Klobutcher

Biological Sciences Faculty Publications

After mating, hypotrichous ciliated protozoa transform a set of their micronuclear chromosomes into thousands of short, linear DNA molecules that form the macronuclear genome. To examine micronuclear genome organization in the hypotrich Euplotes crassus, we have analyzed two cloned segments of micronuclear DNA as well as the macronuclear DNA molecules that are derived from them. E. crassus was found to display a number of features characteristic of other hypotrich genomes, including (i) clustering and close spacing of the precursors of macronuclear DNA molecules, (ii) the frequent occurrence of internal eliminated sequences within macronuclear precursors, (iii) overlapping macronuclear precursors, (iv) lack …

Chromosomal Localization Of Nucleic Acid-Binding Proteins By Affinity Mapping: Assignment Of The Ire-Binding Protein Gene To Human Chromosome 9, Matthias W. Hentze, Hector Seuanez, Stephen J. O'Brien, Joe B. Harford, Richard D. Klausner

Biology Faculty Articles

Three human mRNAs are regulated post-transcriptionally by iron via iron-responsive elements (IREs) contained in each mRNA. A cytoplasmic protein (IRE-BP) binds to these cis-acting elements and mediates the translational regulation of ferritin H- and L-chain mRNA and the iron-dependent stability of transferrin receptor (TfR) mRNA. We have taken advantage of the different mobilities of the human and rodent IRE/IRE-BP complexes on non-denaturing polyacrylamide gels to determine the chromosomal localization of the gene encoding the IRE-BP. Utilizing a panel of 34 different human/rodent hybrid cell lines we have assigned the IRE-BP gene to human chromosome 9. This new technique based on …

The T(11;14)(P15;Q11) In A T-Cell Acute Lymphoblastic Leukemia Cell Line Activates Multiple Transcripts, Including Ttg-1, A Gene Encoding A Potential Zinc Finger Protein, Elizabeth A. Mcguire, Richard D. Hockett, Karen M. Pollock, Marty F. Bartholdi, Stephen J. O'Brien, Stanley J. Korsmeyer

Biology Faculty Articles

Interchromosomal translocations within lymphoid neoplasms frequently involve the antigen receptor genes. We cloned the breakpoints of the t(11;14)(p15;q11) in a CD3-negative T-cell acute lymphoblastic leukemia cell line (RPMI 8402) in order to identify new genes potentially involved in T-cell neoplasia. An extensive comparison of both breakpoints and their germ line counterparts indicated that an inadvertant recombinase-mediated break at chromosome segment 11p15 recombined with the delta T-cell receptor at 14q11. The derivative 11 breakpoint resembles a coding joint in which 11p15 rather than a variable region was introduced 5' to a D_δ1D_δ2J_δ1 intermediate rearrangement. Conversely, the …

Isolation And Partial Characterization Of Basic, Sperm-Specific, Nuclear Proteins In The Dog And Hampster, Richard William Hall

Biological Sciences Theses & Dissertations

The purpose was to determine whether dogs and hamsters contain the same sperm-specific protamine and testis specific proteins (TP) as other eutherian mammals previously studied. The proteins (protamine and TPs) were isolated from spermatids and mature spermatozoa, and were subjected to electrophoresis in an acid-urea, polyacrylamide gel system according to the procedure of Panyim and Chalkley (1962). The results indicated that, regardless of the anatomical location of sperm acquisition, acid precipitation technique, or denaturing agents used, epididymal and testicular samples from both the dog and hamster contained protamine in elongated spermatids and mature spermatozoa. The protamines from dogs and hamsters …

Characterization Of Chromosome Fragmentation In Two Protozoans And Identification Of A Candidate Fragmentation Sequence In Euplotes Crassus, Scott Everet Baird, Lawrence A. Klobutcher

Biological Sciences Faculty Publications

Following the sexual cycle, hypotrichous ciliated protozoans fragment a set of their micronuclear chromosomes to generate the thousands of short, linear DNA molecules present in the transcriptionally active macronucleus. We have used a hybrid selection procedure to examine macronuclear DNA molecules for subtelomeric length heterogeneity to determine whether chromosome fragmentation occurs at unique or multiple sites. The results suggest that multiple, but closely spaced, chromosome fragmentation sites are used by Oxytricha nova. In contrast, Euplotes crassus uses unique chromosome fragmentation sites in a reproducible manner to generate the ends of macronuclear DNA molecules. Additional studies compared DNA sequences in …

Isolation And Immunological Characterization Of A Group Of New B Lymphomas From Cba Mice, Venkatachalam Udhayakumar, Peter H. Brodeur, M. S. Rajagopalan, Stephen Zimmer, Karen E. Pollok, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

We have isolated and characterized a new series of B lymphomas which occurred spontaneously in a group of CBA N mice that were transferred with spleen or lymph node cells from 24-month-old CBA Ca mice. Tumor cell lines from six CBA N mice that received spleen cells were rescued and designated as BKS-2, BKS-3, BKS-4, BKS-5, BKS-6, and BKS-7. Also, tumor cells from a recipient of lymph node cells were rescued and the resulting cell line was designated BKL. These tumor cells expressed membrane immunoglobulin (μ,κ), major histocompatibility complex Class I and Class II molecules, B220, Lyb8, Fc receptors, J11d, …

A Specific Defect In The Proliferative Capacity Of B Cells From Old Mice Stimulated With Autoreactive T Cells, Prakash S. Nagarkatti, Aruna Seth, Mitzi Nagarkatti, Natarajan Muthusamy, Barbara Rychlik, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

B lymphocytes from aged mice were found to be defective in their ability to proliferate in response to stimulation with an autoreactive T cell clone D1.4. The differentiative response leading to antibody secretion was also impaired in the auto D1.4 T cell-stimulated B cells from old mice in comparison to similarly stimulated B cells from young mice. The B cells from old mice were competent in activating the autoreactive T cells such that the T cells were induced to proliferate. The B cell defect appears to be restricted to a certain phase of B cell activation, since old mouse B …

Population Amalgamation And Genetic Variation: Observations On Artificially Agglomerated Tribal Populations Of Central And South America, R Chakraborty, P E Smouse, J V Neel

Faculty, Staff and Student Publications

The interpretation of data on genetic variation with regard to the relative roles of different evolutionary factors that produce and maintain genetic variation depends critically on our assumptions concerning effective population size and the level of migration between neighboring populations. In humans, recent population growth and movements of specific ethnic groups across wide geographic areas mean that any theory based on assumptions of constant population size and absence of substructure is generally untenable. We examine the effects of population subdivision on the pattern of protein genetic variation in a total sample drawn from an artificial agglomerate of 12 tribal populations …

The Gli-Kruppel Family Of Human Genes, John M. Ruppert, Kenneth W. Kinzler, Albert J. Wong, Sandra H. Bigner, Fa-Ten Kao, Martha L. Law, Hector Seuanez, Stephen J. O'Brien, Bert Vogelstein

Biology Faculty Articles

Previous characterization of GLI, a gene found to be amplified and expressed in a subset of human brain tumors, revealed the presence of five tandem zinc fingers related to those of Krüppel (Kr), a Drosophila segmentation gene of the gap class. We have used the GLI cDNA as a molecular probe to isolate related sequences from the human genome. Partial characterization of six related loci, including sequence determination, expression studies, and chromosome localization, revealed that each locus could encode a separate finger protein. The predicted proteins all had similar H-C links, i.e., a conserved stretch of 9 …

Parentage Analysis With Genetic Markers In Natural Populations I The Expected Proportion Of Offspring With Unambiguous Paternity, R Chakraborty, T R Meagher, P E Smouse

Faculty, Staff and Student Publications

Recent studies indicate that polymorphic genetic markers are potentially helpful in resolving genealogical relationships among individuals in a natural population. Genetic data provide opportunities for paternity exclusion when genotypic incompatibilities are observed among individuals, and the present investigation examines the resolving power of genetic markers in unambiguous positive determination of paternity. Under the assumption that the mother for each offspring in a population is unambiguously known, an analytical expression for the fraction of males excluded from paternity is derived for the case where males and females may be derived from two different gene pools. This theoretical formulation can also be …

The Κ-Deleting Element: Germline And Rearranged, Duplicated And Dispersed Forms, Winfried B. Graninger, Paula L. Goldman, Cynthia C. Morton, Stephen J. O'Brien, Stanley J. Korsmeyer

Biology Faculty Articles

Human light chain genes are used in a κ before λ order. Accompanying this hierarchy is the rearrangement of a κ-deleting element (Kde) which eliminates the kappa locus before λ gene rearrangement. In approximately 60% of rearrangements the Kde recombines at a conserved heptamer within the J_κ-C_κ intron. We demonstrated that aberrant V/J rearrangements possessing apparent "N" nucleotides existed 5' to the J_κ-Kde rearrangements. This suggests that the Kde may selectively eliminate nonfunctional V/J alleles. A κ-producing cell that displayed the unusual finding of λ gene rearrangement demonstrated a rearranged Kde. This rearrangement was a …

Properties Of Anti-Lyb-2-Mediated B-Cell Activation And The Relationship Between Lyb-2 Molecules And Receptors For B-Cell Stimulatory Factor-1 On Murine B Lymphocytes, Subbarao Bondada, Joan Morris, A. R. Baluyut

Microbiology, Immunology, and Molecular Genetics Faculty Publications

The murine B-cell differentiation antigen Lyb-2 has been shown to be involved in B-lymphocyte activation and has been postulated by some to be related to a receptor for B-cell stimulatory factor I (BSF-1) (H. Yakura et al., J. Immunol. 137, 1475, 1986). Here we have demonstrated that monoclonal antibody (mAB) to Lyb-2 resembles BSF-1 in its ability to activate small resting B cells and enhancement of surface Ia. Anti-Lyb-2 antibodies bound B cells with very high avidity and were able to induce mobilization of cytosolic-free calcium. Anti-Lyb-2 mAB differs from BSF-1 in that BSF-1 but not anti-Lyb-2 is able to …

Impaired Autoreactive T Cell-Induced T Cell-T Cell Interaction In Aged Mice, Venkatachalam Udhayakumar, Subbarao Bondada, Aruna Seth, Mitzi Nagarkatti, Prakash S. Nagarkatti

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Self-Ia-reactive (autoreactive) L3T4⁺ T cell clones have been shown earlier to stimulate the proliferation of syngeneic naive L3T4⁺ T cells and initiate a T cell-T cell (T-T) interaction leading to the generation of immunoregulatory circuits. Since aging has been shown to be associated with a decline of the immune responsiveness, age-related alterations in the T-T interaction was investigated in the present study. Using several I-E^d-specific autoreactive T cell clones isolated from 2- to 3-month-old (young) DBA/2 mice as stimulators, it was observed that L3T4⁺ T cells from 22- to 24-month-old (aged) DBA/2 mice, failed to …

Isolation Of A Cdna Clone For The Human Laminin-B1 Chain And Its Gene Localization, Michael Jaye, William Modi, George A. Ricca, Robert Mudd, Inc-Ming Chiu, Stephen J. O'Brien, William N. Drohan

Biology Faculty Articles

A cDNA clone encoding the B1 chain of human laminin has been isolated from a human endothelial cell cDNA library. With use of this probe and a panel of rodent/human somatic-cell hybrids and in situ hybridization, the gene for the human laminin-B1 chain has been localized to chromosome 7, band q31.

A Novel Form Of Human Polymorphism Involving The Hdhfr-Ψ1 Pseudogene Identifies Three Rflps, Nicholas P. Anagnou, S. E. Antonarakis, Stephen J. O'Brien, Arthur W. Nienhuis

Biology Faculty Articles

No abstract provided.

Herd Improvement Service Forges Ahead, M P. Bond

Journal of the Department of Agriculture, Western Australia, Series 4

Since its establishment three years ago, the Herd Improvement Service has steadily expanded the range of services provided to Western Australian livestock producers. The genetic advancement and improved hers management offered by the service give farmers the ability to use modern technology in the most cost-effective way. There is an increasing realisation amongst farmers that to survive and remain competitive, it is worthwhile to invest in such technology.

The Herd Improvement Service is a statutory corporation which was formed to improve the productivity of the State's livestock enterprises. Most of the organisation's activities have been directed to servicing the dairy …

Variant Forms Of Ataxia Telangiectasia, A. M. R. Taylor, E. Flude, B. Laher, Michael W. Stacey, E. Mckay, J. Watt, S. H. Greens, A. E. Harding

Bioelectrics Publications

Two ataxia telangiectasia patients with unusual clinical and cellular features are described. Cultured fibroblasts and PHA stimulated lymphocytes from these two patients showed a smaller increase of radiosensitivity than cells from other A-T patients, as measured by colony forming ability or induced chromosome damage respectively, after exposure to ionising radiation. The response of DNA synthesis to irradiation of these cells was, however, the same as for other A-T patients. Cells from a third patient with some clinical features of A-T but with a very protracted course also showed low levels of radiation induced chromosome damage, but colony forming ability and …

Early Events In B-Cell Activation: Anti-Lyb2, But Not Bsf-1, Induces A Phosphatidylinositol Response In Murine B Cells, Stephan A. Grupp, Judith A. K. Harmony, Arthur R. Baluyut, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Previous studies have indicated that the murine surface antigen Lyb2 is involved in an activation pathway that apparently does not involve the surface immunoglobulin receptor. As sIg has been shown to transduce its activation signal through the breakdown of phosphatidylinositol (PI), and since activation via Lyb2 does not involve sIg, it was of interest to determine if binding to Lyb2 generates a PI response. We have demonstrated that an allele-specific monoclonal antibody to Lyb2 (anti-Lyb2 mab), which has previously been shown to drive B cells into S, also activated PI metabolism in these cells. This activation occurred in a dose-dependent …

Barbara Sanford Oral History, Barbara Sanford

Oral History Collection

Susan Mehrtens' Note:

Barbara Sanford's interview echoes most of what I have heard from many of the staff regarding the Lab's mission and goals. The Lab's ideal size, geographical location--with its pluses and minuses--and the environment in which to do science in the current period--all find echoes on other tapes. There is little of candor, consequence or calculation here. Sanford clearly recognizes the stressful nature of scientific life in these times of federal cutbacks, and, as well, the frustrations of directing a laboratory whose size, complexity and needs preclude personal scientific work by the Director. In the face of such …

Donald Bailey Oral History, Donald Bailey

Oral History Collection

Susan Metrhens' Note:

Regarded by many of his peers at Jax as one of the most intuitive, prescient and creative scientists at the Lab, Bailey was a very reflective narrator. While he provides one anecdote of c.c. Little1s sartorial habits, this tape is largely devoid of anecdotal material, and is more a reminiscence by a scientist who was at Jax in the early '50's, who left and returned in 1967. I tried, through a variety of questions, to probe the synergy between Bailey and the Lab, the degree to which his prescience and creativity might be due to the freedom …

Priscilla "Skippy" Lane Oral History, Skippy Lane

Oral History Collection

Susan Mehrtens' Note:

We have in this tape the recollections of one of the most prolific authors in the field of mouse mutants, an indefatigable co-worker of Margaret Dickey, Margaret Green and currently, Muriel Davison. Full of scientific terms, this tape reveals clearly Lane's thorough absorption in the field to which she has devoted nearly the last forty years of her life. Toward the end of this tape, Lane alludes to Earl Green's sexist attitude, which other women have also mentioned to me on and off tape. Lane also waxes eloquently here about her efforts to establish a hiera'rchy within …

Genetic Characterization Of Human C-Rel Sequences, Elise Brownell, Stephen J. O'Brien, William Nash, Nancy Rice

Biology Faculty Articles

We isolated and sequenced a human genomic-DNA segment that is homologous to a portion of v-rel, the transforming gene of reticuloendotheliosis virus (strain T). We also localized the human rel sequences to human chromosome 2 by screening a panel of rodent x human somatic-cell hybrids with the newly described human rel segment.

Thomas Roderick Oral History, Thomas Roderick

Oral History Collection

Susan Metrhens' Note:

Roderick has been a scientist at Jax for thirty years, working in no other place. He was hired just as Earl Green replaced C.C. Little and Roderick worked with and for Green for some dozen years before moving into his own lab, on his own grants. Roderick is both perceptive and analytical and this tape is valuable for his willingness to reflect on trends and personalities. He also has a philosopher's willingness to consider the values that drive the lab. Roderick was forthcoming with anecdotes, including a most telling one of Rich Prehn showing the courage of …

The Effects Of Strain Differences On The Binding Of Concavalian A To Mouse Sperm, Lisa A. Mckay

Honors Theses

The current study, lectins were used as genetic probes to determine the phenotype of the sperm from different strains of mice. Lectins are proteins of non-immune origin, which bind to specific carbohydrates (McCoy, 1986). Being that virtually all cell membranes contain carbohydrate components, lectins have been used to identify all types of cells. In this study, a biotinylated lectin Concavalian A (Con A) derived from Canavalia ensiformis, was used and binding was detected using an avidin-biotin peroxidase (ABP) complex technique. The object of the current research was to 1) determine if mouse strain differences have an effect on lectin binding, …

The Recombinant Dna Controversy: A Contemporary Cautionary Tale, Nina V. Fedoroff

Syracuse Scholar (1979-1991)

A discussion of the scientific and political aspects of recombinant DNA research.

Twenty-Seven Protein Polymorphisms By Two-Dimensional Electrophoresis Of Serum, Erythrocytes, And Fibroblasts In Two Pedigrees, David Goldman, Lynn R. Goldin, P. Rathnagiri, Stephen J. O'Brien, Janice A. Egeland, Carl R. Merril

Biology Faculty Articles

Twenty-seven independent polymorphic loci were detected by two-dimensional electrophoresis (2DE) of serum, erythrocytes, and fibroblasts in two large families and analyzed for linkage to classical genetic markers. We detected seven serum, four erythrocyte, and 17 fibroblast protein loci that exhibited charge variation in these two families and in a sample of unrelated individuals. The genetic basis of protein variants was confirmed by quantitative gene-dosage dependence and by conformance to Mendelian transmission in the two families, except for four rare variants for which transmission analysis was not possible. Linkage analysis demonstrated that each of the variants represent products of independent loci, …

Genetics And Epidemiology Of Gallbladder Disease In New World Native Peoples, K M Weiss, R E Ferrell, C L Hanis, P N Styne

Faculty, Staff and Student Publications

Native peoples of the New World, including Amerindians and admixed Latin Americans such as Mexican-Americans, are highly susceptible to diseases of the gallbladder. These include cholesterol cholelithiasis (gallstones) and its complications, as well as cancer of the gallbladder. Although there is clearly some necessary dietary or other environmental risk factor involved, the pattern of disease prevalence is geographically associated with the distribution of genes of aboriginal Amerindian origin, and levels of risk generally correspond to the degree of Amerindian admixture. This pattern differs from that generally associated with Westernization, which suggests a gene-environment interaction, and that within an admixed population …