Linkage of the locus for conversion of albumin (Acf-1)in the house mouse,Mus musculus

The linkage of the locus for conversion of albumin (Acf-1) has been established on chromosome 1 with the following gene order and recombination percentages: Id-1 19.3±5.2% Acf-1 4.2±1.7% Dip-1 18.4±4.2% Lp.This work was supported by NIH Postdoctoral Fellowship 1F32 GM0527701, Grant BMS75-03397 from the National Science Foundation, Grant ACS VC-17-R from the American Cancer Society, and Contract NO1-ES42159 from the National Institute of Environmental Health Sciences. The Jackson Laboratory is fully accredited by the American Association for the Accreditation of Laboratory Animal Care.     

Mapping of nucleoside phosphorylase (Np-1) and esterase 10 (Es-10) on mouse chromosome 14

A method for detecting two alleles at Np-1 (nucleoside phosphorylase) and three alleles at Es-10 (esterase 10) from mouse blood by cellulose acetate electrophoresis is described. The allelic constitution at these loci for 44 inbred strains and stocks was determined. The location of Np-1 on chromosome 14 was established by backcross experiments in which alleles at Np-1 and Robertsonian translocations were segregating. Es-10 was shown to be linked to Np-1, and the following genetic map of Chr 14 was constructed: centromere-(8.9±4.0 cM)-[Np-1, Wc]-(10.2±1.9 cM)-Es-10-(15.5±3.7 cM)-s. The homologous human loci, NP and ES-D, are not linked.This work was supported by Contract E(11-1)-3267 with the Energy Research and Development Administration, by Contracts NO1-ES4-2156 and NO1-ES4-2159 with the National Institute of Environmental Health Sciences, and by Grants GM 19656 and GM 20919 from the National Institute of General Medical Sciences. D. A. K. was a participant in the 1975 Summer Program for College, Graduate, and Medical Students, which was supported, in part, by the Clark Foundation. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Chromosomal location of soluble glutamic-pyruvic transaminase-1 (Gpt-1) in the mouse

Three alleles at the Gpt-1 (glutamic-pyruvic transaminase-1) locus in the mouse, as identified by electrophoresis on cellulose acetate, and their distribution among inbred mouse strains and wild stocks are described. The Gpt-1 locus was shown to control the soluble form of the enzyme. Three-point linkage analysis established the location of Gpt-1 on chromosome 15 between uw and bt. In addition, a new staining procedure is described that allows the visualization of GPT activity on gels by the deposition of formazan. This is an improvement over previous methods that produced bands of nonfluorescence against a fluorescent background.This investigation was supported in part by Research Grant GM 20919 from the National Institute of General Medical Sciences, and by contract NO1-ES-4-2159 with the National Institute of Environmental Health Sciences. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Histidine decarboxylase phenotypes of inbred mouse strains: A regulatory locus (Hdc) determines kidney enzyme concentration

Mouse kidney histidine decarboxylase (HDC) provides a model system to study genetic control of a hormone-regulated enzyme (inducible by estrogen and thyroxine; repressible by testosterone). Five major HDC phenotypes scored on the basis of (i) enzyme activity and (ii) the difference in activity between the sexes (females usually higher than males) have been discovered by screening 38 strains of mice. One genetic difference between high-activity strains (DBA/2 and C3H/He) and low-activity strains (C57BL/6 and C57BL/10) has been examined in detail. The phenotypic difference segregates as a single gene in both conventional crosses and between recombinant inbred (RI) strains. Immunoprecipitation has shown that the activity difference is due to an alteration in the number of enzyme molecules. The phenotypic difference between high and low strains can therefore be attributed to different alleles of a single regulatory locus, Hdc; the alleleHdc ^d determines low HDC concentration, and the allele Hdc ^d high concentration. Hdc has been mapped to chromosome 2 using data from both comparisons of strain distribution patterns of previously mapped loci within RI strains and a conventional three-point cross. The probable gene order is B2m-pa-Hdc, with map distances of 3.1±1.7 and 2.0±1.4 cM, respectively.This work was supported by an MRC project grant to Grahame Bulfield, an SERC research studentship to S. A. M. Martin, and NIH Research Grant GM 18684 from the National Institute of General Medical Sciences to B. A. Taylor. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory animal care.     

Evolution of mammalian carbonic anhydrase loci by tandem duplication: Close linkage of Car-1 and Car-2 to the centromere region of chromosome 3 of the mouse

Electrophoretic variants of two carbonic anhydrase enzymes, CAR-1 (CA I) and CAR-2 (CA II), have been found in the laboratory mouse, Mus musculus. These two loci are closely linked to each other and are located on chromosome 3 near its centromere. The close linkage of Car-1 and Car-2 supports the hypothesis that the present-day carbonic anhydrase loci are the result of tandem duplication of an earlier carbonic anhydrase locus with subsequent divergence. The red blood cells of mice of the subspecies M. m. casteneus have significantly reduced levels of CAR-1 and CAR-2.This research was supported in part by Research Grants GM-20919 from the National Institute of General Medical Sciences and CA-01074 from the National Cancer Institute, and by Contracts E(11-1)-3267 with the Energy Research and Development Administration and NO1-ES-4-2159 with the National Institute of Environmental Health Sciences. The Jackson Laboratory is fully certified by the American Association for Accreditation of Laboratory Animal Care.     

Evolution of 5S ribosomal RNA genes in the chromosomes of the virilis group of Drosophila

Drosophila melanogaster 5S ribosomal RNA labeled with ¹²⁵I was used as an in situ hybridization probe to localize complementary sequences in chromosomes of species in the Drosophila virilis group. Whereas virilis, the ancestral species, has two different 5S gene loci, the derived species show only one of these loci; in the two lines that have evolved from virilis it is the opposite locus that is conserved. The possible events leading to such an arrangement are discussed.The author was a Predoctoral Fellow supported by Grant GM 1974 from the National Institute of General Medical Sciences, National Institutes of Health.Contribution from Oak Ridge National Laboratory, operated by the Union Carbide Corporation for the U.S. Energy Research and Development Administration.     

Linkage analyses using biochemical variants in mice. II. Levulinate dehydratase and autosomal glucose 6-phosphate dehydrogenase

The level of hepatic -aminolevulinate dehydratase varies among inbred strains of mice and is regulated by codominant alleles at the Lv locus. Twenty-two inbred strains have been classified with respect to this locus. Lv is 5±2 recombination units from brown, b, in linkage group VIII. The locus for autosomal glucose 6-phosphate dehydrogenase (Gpd-1) has also been assigned to linkage group VIII and is 32±5 units from brown. The order of the loci is Lv-b-Gpd-1. Incidental note is made of linkage between the malic dehydrogenase (Mdh-1) and dilute (d) loci, linkage group II, with 10±3 % recombination between the two.Supported by the Roche Institute of Molecular Biology, Nutley, New Jersey, and by Public Health Service Research Grant CA-05873 from the National Cancer Institute.     

Linkage of isozyme loci in the mouse: Phosphoglucomutase-2 (Pgm-2), mitochondrial NADP malate dehydrogenase (Mod-2), and dipeptidase-1 (Dip-1)

The linkages of the isozyme genes Mod-2, Pgm-2, and Dip-1 have been determined in tests with established linkage group markers among inbred strains of mice. Unique alleles for both Mod-2 and Pgm-2 have been observed in the strain of SM/J. Linkage was determined from backcross progeny of the matings C57BL/6J×(SM/J×C57BL/6J)F₁, (SM/J×SWR/J)F₁×SM/J, and (SM/J×SWR/J)F₁×SJL/J. The gene Mod-2 is on linkage group 1. In a three-point cross of the loci Gpi-1, c, and Mod-2, the c locus was determined to be the middle gene. No double crossovers were observed. Our combined data show the following linkages: Gpi-1 to c, 28.3±3.2%; Gpi-1 to Mod-2, 33.3±3.0%; and c to Mod-2, 4.1±2.8%. The proposed gene order for four markers on LG I is Gpi-1-p-c-Mod-2. The gene Pgm-2 was linked to Gpd-1 (27.0±4.2%) on LGVIII. Two backcrosses segregating for Pgm-2 and b, (SM/J×DBA/2J) F₁×DBA/2J and (SM/J×DBA/2J)F₁×C57BR/cdJ, showed 9.1±4.3% recombination. The proposed gene order on LG VIII is b-Pgm-2-Gpd-1. The genes Pgm-1 and Pgm-2 are not linked (53.4±4.4%). Linkage of the isozyme genes Dip-1 and Id-1 on LG XIII was observed in backcross progeny of the crosses (SJL/J×C57BL/6J)F₁×SJL/J and C57BL/6J×(SM/J×C57BL/6J)F₁. The combined recombination was 23.8±2.8%. Two cases are established where genes whose enzyme products share substrate affinities (Pgm-1 and Pgm-2; Mod-1 and Mod-2) are not linked. Our data generally support the conclusion that functionally or metabolically related isozyme genes are not contiguous on mouse linkage groups.This investigation was supported in part by Public Health Service General Research Support Grant GM-09966 and in part by Public Health Service Training Grant 5T01 HD-00032-07 from the National Institute of Child Health and Human Development, and by Atomic Energy Commission contract AT(30-1)-3671.     

Murine liver arylsulfatase B processing influenced by region on chromosome 17

SM/J liver arylsulfatase B has a more rapid electrophoretic mobility and occurs as a series of more acidic isozymes following electrofocusing in narrow pH gradients than the liver enzyme from C57BL/6J mice. The SM/J and C57BL/6J electrofocusing patterns were both converted to a single isozyme with similar isoelectric points by pretreatment with neuraminidase, suggesting that the SM/J and C57BL/6J isozymes differed with respect to their sialic acid content. Arylsulfatase B electrofocusing and thermostability phenotypes segregated independently among progeny of SM/J×C57BL/6J crosses, suggesting that the electrofocusing phenotypes were not determined by different alleles at As-1, the putative structural locus for arylsulfatase B. Comparison of the joint segregation of hepatic acid phosphatase electrophoretic patterns and liver arylsulfatase B electrofocusing profiles revealed that the electrofocusing profiles may be determined by a region on chromosome 17 near or identical to Apl. Kidney, brain, and spleen arylsulfatase B electrofocusing patterns did not appear to differ between SM/J and C57BL/6J mice.This research was supported in part by Biomedical Sciences Research Support Grant RR-07030, by NIGMS Grant 1-RO1GM27707-01, and by Grant 1–570 from the National Foundation/March of Dimes.     

Comparative analysis of malate dehydrogenases of Drosophila melanogaster

The malate dehydrogenases of D. melanogaster have been resolved into a cytoplasmic form (cMDH) and a mitochondrial matrix form (mMDH). Flies homozygous for allozyme variants exhibit isozymes of cMDH detected by starch gel electrophoresis and acrylamide gel isoelectric focusing. The basis of these isozymes was investigated, and the results suggest either conformational or epigenetic modification of isozymes. The probable structural gene for cMDH (Mdh-1) has been mapped genetically by allozyme variants to II-35 ± 3 and cytologically by monitoring gene dosage in segmental aneuploids to between 28D and 29F on II-L of the Drosophila salivary gland chromosome map. The structural gene for mMDH is neither identical to nor in the near chromosomal proximity of Mdh-1. Nevertheless, the two enzymes exhibit markedly similar properties with respect to (1) catalytic activity, (2) pH optima, (3) pH optimum shift in response to different ionic environments, and (4) molecular weight as determined by sucrose density gradient sedimentation.This project was supported by NIH postdoctoral research fellowship No. 6-FO2-GM-49, 633-01 from the National Institute of General Medical Sciences.     

Phenol oxidase activity and the Lozenge locus of Drosophila melanogaster

We have found that the phenol oxidase activity in 50-hr Drosophila melanogaster pupae is much greater than that of adult flies. The mutants lz and lz ^g have all of the phenol oxidase components present in wild type, whereas the mutant tyr-1 has all of the wild-type components but the activity of each component is greatly reduced in comparison with wild-type activity. The newly discovered lozenge allele, lz ^rfg, lacks all phenol oxidase activity.Predoctoral fellow supported by Grant GM 1974 from the National Institute of General Medical Sciences, National Institutes of Health.The Oak Ridge National Laboratory is operated for the U.S. Atomic Energy Commission by Union Carbide Corporation.     

An acid phosphatase locus expressed in mouse kidney (Apk) and its genetic location on chromosome 10

A genetic locus controlling the electrophoretic mobility of an acid phosphatase in mouse kidney is described. This locus, called acid phosphatase-kidney (Apk), is not expressed in erythrocytes, liver, spleen, heart, lung, brain, skeletal muscle, stomach, or testes. The product of Apk hydrolyzes the substrate naphthol AS-MX phosphoric acid but is not active on a-naphthylphosphate or 4-methylumbelliferylphosphate. It is not inactivated by 50 C for 1 hr, nor is its electrophoretic mobility altered by incubation with neuraminidase. The locus is invariant among 31 inbred strains (Apk ^a), with a variant allele (Apk ^m) observed only in Mus musculus molossinus. Codominant expression was observed in F₁ hybrids of M. m. molossinus and inbred strains. Apk was mapped on Chr 10, near the neurological mutant waltzer (v).This work was supported by Contract NO1-ES42159 from the National Institute of Environmental Health Sciences and by Grant 1-476 from the National Foundation—March of Dimes. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Metabolic properties of erythrocytes of normal and genetically anemic mice

The murine hemolytic anemias microcytosis (gene symbol mk), normoblastosis (nb), spherocytosis (sph), and hemolytic (ha) are inherited as autosomal recessive diseases and resemble the human hereditary hemolytic anemias caused by defective enzyme activities in erythrocytes. The activities of 14 different enzymes of the glycolytic and pentose phosphate pathways were compared in erythrocytes from normal and anemic mice, but no quantitative differences suggesting enzyme deficiency were found. There were no major changes in reduced glutathione, NAD, NADP, or methemoglobin content. The rate of entry of glucose into the glycolytic and hexose monophosphate shunt pathways of intact erythrocytes was higher in mk/mk erythrocytes than predicted. Interpretation of studies of erythrocytes from anemic mice is generally complicated by the extremely high reticulocyte and nucleated cell counts in ha/ha, sph/sph, and nb/nb mice.Investigations in Kentucky (Dr. Hutton) were supported by Research Career Development Award 1-K4-AM-70, 186-01 and NIH Research Grant AM 16013-01 from the National Institute of Arthritis and Metabolic Diseases, and those at The Jackson Laboratory (Dr. Bernstein) by NIH Research Grant HD-00254 from the National Institute of Child Health and Human Development, by U.S. Atomic Energy Commission Contract AT(30-1)-1800, and in part by the George W. Perkins Memorial Fund and by income from the Endowment Funds of The Jackson Laboratory. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care.     

Regulation of gene function: A comparison of enzyme activity levels in relation to gene dosage in diploids and triploids of Drosophila melanogaster

A study of gene activity in diploid and triploid Drosophila melanogaster females has been performed. Levels of enzyme activity of X-linked glucose 6-phosphate and 6-phosphogluconate dehydrogenases and autosome-linked -glycerophosphate and NADP-dependent isocitrate dehydrogenases were measured and correlated with DNA content, in crude extracts of whole flies or thoraces. The results show that the contribution of each dose of a given gene to the level of enzyme activity is equal in diploid and triploid cells. These findings are discussed in the context of the phenomenon of dosage compensation.This investigation was supported by Research Grant GM-15691 and Genetics Training Grant 2 T1-GM-685 of the National Institutes of Health.Recipient of a Research Career Development Award (K4-GM-13, 277) from the National Institute of General Medical Sciences.     

The development and evaluation of immunodiffusion,immunofluorescence and physiological tests for the differentiation of atypical trichophyton rubrum and T. mentagrophytes isolates

Five laboratory procedures: 1) immunodiffusion, 2) immunofluorescence, 3) in vitro hair perforation, 4) pigment stimulation, and 5) a urease test were compared for their ability to differentiateT. rubrum fromT. mentagrophytes. Of the physiological tests, thein vitro hair perforation technique was the most reliable for differentiating the two species. With the serological tests, the organisms were not differentiated by immunodiffusion, but if appropriate dilutions of the conjugates were used in immunofluorescence testing, most isolates could be differentiated.A portion of a Dissertation submitted by the senior author to the University of North Carolina in partial fulfillment of the requirements for the degree of Doctor of Public Health in the School of Public Health. Training was provided by the Laboratory Director's Program which is supported by Training Grant TO1 GM 00567-07 from the National Institute of General Medical Sciences, National Institutes of Health, United States Public Health Service. The laboratory research was performed at the Laboratory Division, Center for Disease Control, under the supervision of William Kaplan.     

Ectopic pairing and evolution of 5S ribosomal RNA genes in the chromosomes of Drosophila funebris

5S ribosomal RNA from Drosophila melanogaster labeled with ¹²⁵I was used to locate the 5S rRNA genes in chromosomes of D. funebris by means of in situ hybridization. Silver grains were observed at three distinct sites, one of which was a recognized reverse repeat. Only one half of the reverse repeat, however, hybridizes with 5S rRNA and the significance of this phenomenon is discussed. A case of ectopic pairing between two different 5S sites in the genome is reported, and the significance of ectopic pairing is considered.The author was a Predoctoral Fellow supported by Grant GM 1974 from the National Institute of General Medical Sciences, National Institutes of Health.Contribution from Oak Ridge National Laboratory, operated by the Union Carbide Corporation for the U.S. Energy Research and Development Administration.     

Linkage relationships of biochemical markers to Q- and C-band variants in a large black kindred

Summary Lod scores are reported for 86 biochemical to cytogenetic marker comparisons in a Black kindred. Analysis with unconfirmed locus assignments resulted in 12 exclusions of close linkage.This paper is based on a thesis submitted by the senior author in partial fulfillment of the Ph.D. degree at North Carolina State University, Raleigh.Research supported by the National Heart and Lung Institute Grant HLO-3341, National Institute of Mental Health Grant MH 26621, and National Institute of General Medical Sciences Grant GN 16697; paper number 6281 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina.     

Location of phosphorylase kinase (Phk) in the mouse X chromosome

The phosphorylase kinase deficiency (Phk) locus has been located in the mouse X chromosome, the order of genes being centromere-Bn-Phk-Ta-jp. Since the Phk locus of the mouse may be identical to the locus responsible for the X-linked phosphorylase kinase deficiency trait of man, and there may be a high degree of gene-order homology in the X chromosome of all mammals, the location of Phk in the mouse reported here may aid in locating the phosphorylase kinase gene on the X chromosome of man.This research was supported by grants AM 13359 (to F.H.) and AM 14461 (to D.L.C.) from the National Institute of Arthritis and Metabolic Diseases, and by an allocation (to E.M.E.) from NIH General Research Support Grant RR-05545 from the Division of Research Resources to The Jackson Laboratory. F.H. is a recipient of a Research Career Development Award (AM 46 421) of the National Institute of Arthritis and Metabolic Diseases.     

In vitro translation of maize ADH: Evidence for the anaerobic induction of mRNA

Total cellular RNA from anaerobically stressed maize seedling roots was used to stimulate in vitro translation of authentic maize alcohol dehydrogenase (ADH) in a rabbit reticulocyte lysate system. Total products from such reactions were displayed on NEPHGE-SDS two-dimensional gels and the Adhl-specific translation products were identified by using RNA from sib seedlings segregating for Adhl charge and size variants. The application of a rapid RNA isolation procedure allowed the efficient isolation of biologically active RNAs from small amounts of seedling material. Maize ADHs translated in vitro are identical in size to in vivo ADH. Further, no ADH was detected in the products of an in vitro translation reaction stimulated by total RNA from aerobically grown seedlings. This suggests that induction of ADH protein by anaerobic stress is accomplished by production of Adh mRNA rather than activation of sequestered mRNA. The mRNAs for maize ADH1 and ADH2 are among a small class of mRNAs induced during anaerobiosis.Research was supported by NSF Grant PCM 76-11009. M.D.B. is supported by National Institute of Health Grant PHS 5 T32 GM07227-04. R.J.F. is a Predoctoral Trainee in Genetics supported by National Institute of Health Training Grants 82 and 7757 from the National Institute of General Medical Sciences.     

Chromosomal location of the catalase structural genes in Zea mays using B-A translocations

Summary B-A translocations have been used to map the catalase genes, Cat1, Cat2, and Cat3 of Zea mays. Cat1 was found to be on the short arm of chromosome 5, 9.1 map units from brittle endosperm (bt ₁). Cat2 was located on chromosome 1S, while Cat3 was located on the distal half of chromosome 1L. There was no linkage between Cat2 and Cat3. The significance of mapping the catalase structural genes is discussed.This research was supported by Grant No. GM22733 from the National Institute of General Medical Sciences, National Institutes of Health, USPHS to JGS.This is Paper No. 6437 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC