Report of the Second International Workshop on Y Chromosome Mapping 1995.
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brt encodes a receptor-type protein-tyrosine kinase and was isolated from fetal mouse brain by using a PCR-mediated cloning procedure; this gene is expressed preferentially in the brain of both embryo and adult. Chromosomal locations of the mouse and rat brt genes were determined by fluorescence in situ hybridization using a mouse cDNA fragment as a probe. The brt gene was localized to mouse Chromosome 2F1 and rat Chromosome 3q36.1, where a conserved linkage homology has been identified between the two species. Interspecific backcross analysis genetically demonstrated no recombination between the mouse BRT locus and D2Mit63.
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The neural-specific calmodulin-sensitive adenylyl cyclase (type I), which was first cloned from bovine brain, has been implicated in learning and memory. The objective of this study was to clone and determine the chromosomal localization of human fetal brain type I adenylyl cyclase. A 3.8-kb cDNA clone was isolated that contained sequence coinciding with the 3' end 2553 nucleotides of the bovine open reading frame. This clone shows 87% nucleotide and 92% translated amino acid sequence identity to the bovine clone. The most significant sequence differences were in the carboxy-terminal 100 amino acid residues. This region contains one of several possible calmodulin binding domains and the only putative cAMP-dependent protein kinase A phosphorylation site. A chimera was constructed that contained the 5' half of the bovine type I adenylyl cyclase and the 3' half of the human type I adenylyl cyclase. The activity of the chimeric gene product and its sensitivity to calmodulin and calcium were indistinguishable from those of the bovine type I adenylyl cyclase. In situ hybridization was used to localize the human type I adenylyl cyclase gene to the proximal portion of the short arm of chromosome 7.
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The cDNAs for human and murine Receptor Activity Modifying Proteins and for the associated murine Calcitonin Receptor Like Receptor were isolated. The human RAMP1 and RAMP3 genes possess two introns and human RAMP2 possesses three introns. Human RAMP1 was assigned to chromosome 2q36-->q37.1, RAMP2 to 17q12-->q21.1 and RAMP3 to 7p13-->p12. Mouse Ramp1 was assigned to chromosome 1 and Ramp2 and Ramp3 were assigned to chromosome 11.
Cytochrome P450 17 alpha-hydroxylase/C17-20 lyase (P45017 alpha) catalyzes the conversion of C-21 steroids to C-19 steroids in gonads. A full-length mouse cDNA encoding P450 17 alpha was isolated from a mouse Leydig cell library and characterized by restriction mapping and sequencing. The predicted amino acid sequence has 83% homology to rat, 66% homology to human, and 62% homology to bovine P45017 alpha amino acid sequences. The protein is 507 amino acids in length, which is 1 amino acid shorter than the human protein and 2 amino acids shorter than the bovine protein. The structural gene encoding P450 17 alpha (Cyp17) was localized utilizing an interspecific testcross to mouse chromosome 19, distal to Got-1. Another cytochrome P450, P4502c (Cyp2c), also is located at the distal end of chromosome 19. CYP17, CYP2c, and GOT1 have been mapped to human chromosome 10, with CYP2C and GOT1 mapped to the distal region, q24.3 and q25.3, respectively. The data in the present study indicate conserved syntenic loci on mouse chromosome 19 and human chromosome 10 and predict that the structural gene encoding P45017 alpha will be found distal to GOT1 on human chromosome 10.
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Chromosomal locations of members of the xenotropic-related env gene family in the mouse genome have been determined. Endonuclease restriction site polymorphisms detected by molecular hybridization were used to study the inheritance of mink cell-focus inducing and xenotropic env gene-related sequences in recombinant inbred strains of mice. Some of the endogenous env sequences appear to be closely linked to genes determining leukemia virus induction and to genes involved in the immune response, such as the heavy and light chains of the immunoglobulin molecules or allotypic determinants on B and T lymphocytes. The use of probes that detect restriction fragment length polymorphisms in a small family of dispersed sequences promises to yield a large number of markers that can be used together with recombinant inbred strains for efficient mapping of the mouse genome.
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The validity of mouse liver tumors is controversial in the risk assessment of carcinogenicity of chemicals in humans, because mice used in carcinogenicity bioassays are genetically predisposed to liver tumors. The argument could be resolved once liver tumor susceptibility genes have been cloned and their role in liver tumor development elucidated. We performed a genetic linkage analysis to map murine liver tumor susceptibility genes, as a first step toward their identification. An F2 population of 87 urethane-treated male A/J x C3H/He mice was scored with 83 genetic markers. Three regions, localized on chromosomes 7, 8, and 12, were found to contain putative liver tumor susceptibility genes.
We molecularly characterized the second gene, c-yes-2, of two copies of yes-related genes which we previously found to contain in the human genome. First, nucleotide sequence analysis revealed that the c-yes-2 gene is a pseudogene of the c-yes-1 gene. Second, by using two independent methods, hybridization of both DNAs from sorted chromosomes and metaphase spreads with c-yes-2 DNA, we assigned the c-yes-2 gene to chromosome 22q11.2. This chromosomal localization is consistent with that given in our previous report. The failure of proper mapping in our experiment might have been caused by instability of hybrid cell clones.
Two quantitative trait loci (QTL) affecting susceptibility of the mosquito Aedes aegypti to the filarial worm parasite Brugia malayi were identified using restriction fragment length polymorphism (RFLP) markers. The first locus, fsb[1,LF178], resides within a 10 cM interval on chromosome 1 and exhibits a recessive effect with respect to susceptibility. The second locus, fsb[2,LF98], resides within a 9 cM interval on chromosome 2 and exhibits an additive effect on susceptibility. Significant epistasis was detected between these loci, although the effect of fsb[2,LF98] was dependent on the genetic background of the mosquito strains. Suggestions for a standard QTL nomenclature are included.
Linear IgA disease is characterized by circulating IgA autoantibodies recognizing basement membrane zone components, including an anchoring filament protein, ladinin. In this study, we have cloned the mouse ladinin cDNA, elucidated the intron-exon organization of the corresponding gene (Lad1), and determined its chromosomal assignment. We have also characterized the promoter region of Lad1 and examined its tissue-specific expression. The mouse Lad1 gene consists of 10 exons spanning approximately 13.4 kb of the mouse genome on chromosome 1, and Southern analysis suggested that Lad1 is a single-copy gene. The coding region comprises 1584 nucleotides and encodes a 528-amino-acid polypeptide with a calculated molecular mass of 59 kDa. The deduced polypeptide contained two putative N-glycosylation and two O-glycosylation sites, and sequence analysis predicted a 15-amino-acid signal peptide. The 5' upstream region demonstrated the presence of consensus cis-elements for AP2 and SP1 and was GC rich, consistent with eukaryotic promoter. Northern analysis revealed expression in cultured keratinocytes, but not in fibroblasts, with the mRNA transcript being approximately 2.5 kb in size. A significant level of expression was also noted in the kidney and lung, and to a lesser degree in the liver, spleen, and brain. Ladinin is a novel component of the basement membranes and may function in contributing to the stability of the association of the epithelial layers with the underlying mesenchyme.
The structure of Krox-24, a mouse zinc-finger-encoding gene that is transiently activated during G0/G1 transition, has been established. Krox-24 is located on mouse chromosome 18, bands C-D. The gene product, as anticipated for a putative DNA-binding protein, is localized within the cell nucleus. The Krox-24 5'-flanking region contains a series of serum response elements (SREs) similar to the SRE observed upstream of the c-fos proto-oncogene. These elements can substitute for the c-fos SRE, their effect is cumulative and they bind the same cellular factor, the serum response factor (SRF), as the c-fos SRE. This suggests that the SRE and its cognate protein are likely to be involved in the regulation of Krox-24 and presumably of other immediate-early serum response genes. SRE and SRF therefore constitute key components in the regulatory pathway leading from mitogenic stimulation to cellular proliferation.
Males of the noctuid moths, Heliothis virescens and H. subflexa locate mates based on species-specific responses to female-emitted pheromones that are composed of distinct blends of volatile compounds. We conducted genetic crosses between these two species and used AFLP marker-based mapping of backcross families (H. subflexa direction) to determine which of the 30 autosomes in these moths contained quantitative trait loci (QTL) controlling the proportion of specific chemical components in the pheromone blends. Presence/absence of single H. virescens chromosomes accounted for 7-34% of the phenotypic variation among backcross females in seven pheromone components. For a set of three similar 16-carbon acetates, two H. virescens chromosomes interacted in determining their relative amounts within the pheromone gland and together accounted for 53% of the phenotypic variance. Our results are discussed relative to theories about population genetic processes and biochemical mechanisms involved in the evolution of new sexual communication systems.