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Chromosomal mapping and expression of the human B120 gene.

We previously reported a novel human cDNA, designated B120, containing a CAG repeat length polymorphism and many repeat units, loosely identified as YXQQP which is found in several human RNA binding proteins. In the present study, the B120 gene was mapped to human chromosome 1p35-36.1 by fluorescence in situ hybridization (FISH). Several human disorders, including that of Schnyder crystalline corneal dystrophy, have been mapped to this region by genetic linkage. Schnyder crystalline corneal dystrophy is thought to be a primary abnormality of corneal lipid metabolism, resulting in opacification secondary to lipid accumulation. In order to examine the function of B120, we introduced B120 cDNA with an expression vector into various cell lines including Cos1, C3H/10T1/2 and NIH/3T3 cells. These transfected cells exhibited small cytoplasmic spherical bodies. The cytoplasmic bodies appeared to be fat droplets on electron microscopy and histochemical staining. These findings suggested that B120 gene expression is associated with lipid metabolism, and that overexpression of B120 may result in lipid deposition in various cells, including those of fibroblastic cell lines. Since the cornea is composed of fibroblastic cells, overfunction of B120 could be related to the pathogenesis of Schnyder crystalline corneal dystrophy.

3T3 Cells↗

The mouse aldehyde oxidase gene: molecular cloning, chromosomal mapping and functional characterization of the 5'-flanking region.

In this article, we report on the chromosome mapping and molecular cloning of the genetic locus encoding the mouse molybdo-iron/sulfur-flavoprotein aldehyde oxidase. The aldehyde oxidase locus maps to mouse chromosome 1 band C1-C2, as determined by fluorescence in situ hybridization experiments conducted on metaphase chromosomes. The gene is approximately 83 kb long and consists of 35 exons. The exon/intron boundaries are perfectly conserved relative to the corresponding human homolog and almost completely conserved relative to the mouse xanthine oxidoreductase gene. This further supports the concept that the aldehyde oxidase and xanthine oxidoreductase loci evolved from the same ancestral precursor by a gene duplication event. The position of a major transcription start site was defined by primer extension and RNase mapping analysis. The 5'-flanking region of the mouse aldehyde oxidase gene contains a functional and orientation-dependent promoter as well as several putative binding sites for known cell-specific and general transcription factors. Deletion analysis of the 5'-flanking region defines an approximately 470 bp DNA stretch which is necessary and sufficient for the transcription of the mouse aldehyde oxidase gene.

Aldehyde Oxidase↗

Chromosome map of the thermophilic archaebacterium Thermococcus celer.

A physical map for the chromosome of the thermophilic archaebacterium Thermococcus celer Vu13 has been constructed. Thirty-four restriction endonucleases were tested for their ability to generate large restriction fragments from the chromosome of T. celer. Of these, the enzymes NheI, SpeI, and XbaI yielded the fewest fragments when analyzed by pulsed-field electrophoresis. NheI and SpeI each gave 5 fragments, while XbaI gave 12. The size of the T. celer chromosome was determined from the sum of the apparent sizes of restriction fragments derived from single and double digests by using these enzymes and was found to be 1,890 +/- 27 kilobase pairs. Partial and complete digests allowed the order of all but three small (less than 15 kilobase pairs) fragments to be deduced. These three fragments were assigned positions by using hybridization probes derived from these restriction fragments. The positions of the other fragments were confirmed by using hybridization probes derived in the same manner. The positions of the 5S, 16S, and 23S rRNA genes as well as the 7S RNA gene were located on this map by using cloned portions of these genes as hybridization probes. The 5S rRNA gene was localized 48 to 196 kilobases from the 5' end of the 16S gene. The 7S RNA gene was localized 190 to 504 kilobases from the 3' end of the 23S gene. These analyses demonstrated that the chromosome of T. celer is a single, circular DNA molecule. This is the first such demonstration of the structure of an archaebacterial chromosome.

Archaea↗

Comparative polytene chromosome maps of D. montana and D. virilis.

Chromosomal inversion polymorphism was characterized in Finnish Drosophila montana populations. A total of 14 polymorphic inversions were observed in Finnish D. montana of which nine had not been described before. The number of polymorphic inversions in each chromosome was not significantly different from that expected, assuming equal chance of occurrence in the euchromatic genome. There was, however, no correlation between the number of polymorphic inversions and that of fixed inversions in each chromosome. Therefore, a simple neutral model does not explain the evolutionary dynamics of inversions. Furthermore, in contrast to results obtained by others, no significant correlation was found between the two transposable elements (TEs) Penelope and Ulysses and inversion breakpoints in D. montana. This result suggests that these TEs were not involved in the creation of the polymorphic inversions seen in D. montana. A comparative analysis of D. montana and Drosophila virilis polytene chromosomes 4 and 5 was performed with D. virilis bacteriophage P1 clones, thus completing the comparative studies of the two species.

Animals↗

Chromosome mapping of the murine syndecan gene.

The chromosomal localization of the murine syndecan gene was determined by analysis of DNA from a panel of mouse-hamster cell hybrids containing various mouse chromosomes, detection of immunoreactive syndecan in culture medium of these cells, and linkage analysis of a mouse interspecific backcross. Southern analysis of the mouse-hamster cell hybrid DNA shows two distinct hybridizing sequences, one on mouse Chromosome 12 and the other on the X chromosome. Localization of the syndecan gene to mouse Chromosome 12 was determined by detection of immunoreactive syndecan in the culture medium of cell hybrids containing mouse Chromosome 12. Hybrids containing other mouse chromosomes were negative. Linkage analysis by Southern hybridization of DNA from a mouse interspecific backcross using a syndecan-specific probe localized the syndecan gene locus, Synd, to the proximal end of Chromosome 12, tightly linked to the Pomc-1 and Nmyc loci. The syndecan gene is likely on human Chromosome 2 because this region shows conservation of synteny between mouse and human chromosomes.

Animals↗

Chromosomal mapping of the human annexin IV (ANX4) gene.

Annexin IV (placental anticoagulant protein II) is a member of the annexin or lipocortin family of calcium-dependent phospholipid-binding proteins. A cDNA for human annexin IV was isolated from a placental library that is 675 bases longer in the 3' untranslated region than previously reported, indicating the existence of alternative mRNA processing for this gene. Genomic Southern blotting with a cDNA probe indicated a gene size of 18-56 kb. Primers developed for polymerase chain reaction (PCR) allowed amplification of a 1.6-kb portion of the ANX4 gene. DNA sequence analysis showed that this PCR product contained a single intron with exon-intron boundaries in exactly the same position as in the mouse annexin I and annexin II genes. PCR analysis of a somatic cell hybrid panel mapped the ANX4 gene to chromosome 2, and in situ hybridization with a cDNA probe showed a unique locus for ANX4 at 2p13. This study provides further evidence that genes for the annexins are dispersed throughout the genome but are similar in size and exon-intron organization.

Amino Acid Sequence↗

Molecular cloning, chromosome mapping and characterization of a testis-specific cystatin-like cDNA, cystatin T.

The cystatin superfamily of cysteine proteinase inhibitors consists of three major families. In the present study, we report the cloning of the cDNA for mouse cystatin T, which is related to family 2 cystatins. The deduced amino acid sequence of cystatin T contains regions of significant sequence homology including the four highly conserved cysteine residues in exact alignment with all cystatin family 2 members. However, cystatin T lacks some of the conserved motifs believed to be important for inhibition of cysteine proteinase activity. These characteristics are seen in two other recently cloned genes, CRES and Testatin. Thus, cystatin T appears to be the third member of the CRES/Testatin subgroup of family 2 cystatins. The mouse cystatin T gene was mapped on a region of chromosome 2 that contains a cluster of cystatin genes, including cystatin C and CRES. Northern blot analysis demonstrated that expression of mouse cystatin T is highly restricted to the mouse testis. Thus, a shared characteristic of the cystatin family 2 subgroup members is an expression pattern limited primarily to the male reproductive tract.

Animals↗

Chromosomal mapping, sequence and transcription analysis of the porcine fertilin beta gene (ADAM2).

Fertilin beta (ADAM2) forms a part of the heterodimeric surface protein fertilin, found on the plasma membrane of mammalian sperm, and has been implicated in the process of sperm-egg fusion. Analysis of cDNA products obtained from adult porcine testis mRNA has presented a sequence corresponding to 2620 bp of the ADAM2 gene. This sequence contained an open reading frame encoding a 735-amino acid protein and homologous to ADAM2 genes known in other mammalian species. Polymerase chain reaction (PCR) analysis of genomic DNA showed that the 2620 bp of cDNA sequence comprises at least 21 exons and spans approximately 76 kb of genomic DNA, with its size and structure being relatively conserved between mouse, human and pig. Fluorescence in situ hybridization was used to map ADAM2 to chromosome 15 of the pig, using a bacterial artificial chromosome clone from the PigE BAC library. This finding is consistent with comparative mapping experiments performed between pig and human chromosomes. Analysis of nine mRNA samples, by reverse transcriptase-PCR, from different porcine tissues has also suggested that expression of ADAM2 is limited to the testis, a finding that is consistent with other mammalian species.

ADAM Proteins↗

Chromosome mapping of the human RAS-related RAP1A, RAP1B, and RAP2 genes to chromosomes 1p12----p13, 12q14, and 13q34, respectively.

Three human cDNAs encoding new RAS-related cDNAs, designated RAP1A, RAP1B, and RAP2, have been isolated previously. The encoded proteins are highly related to RAS in the effector region and share an overall identity with RAS of approximately 50%. Using the complete cDNAs or parts thereof as probes, each RAP gene has been localized on human chromosomes by in situ hybridization. The three genes RAP1A, RAP1B, and RAP2 have been assigned to chromosome bands 1p12----p13, 12q14, and 13q34, respectively.

Autoradiography↗

Chromosomal mapping of HSPCB and MYL1 expressed abundantly in the bovine fetus.

Chromosomal mapping of expressed sequence tags for HSPCB and MYL1 expressed abundantly in the bovine fetus was performed by analyzing bovine/murine somatic cell hybrid DNAs with polymerase chain reaction (PCR) using primers specific for those 3'-untranslated regions. HSPCB and MYL1 were assigned to bovine chromosomes 23 and 2, respectively.

Animals↗

Chromosomal mapping of two members of the human glutamate dehydrogenase (GLUD) gene family to chromosomes 10q22.3-q23 and Xq22-q23.

Glutamate dehydrogenase (GLUD) is an important mitochondrial enzyme that participates in neuronal transmission by catalyzing the deamination of L-glutamate, which serves as a potent excitatory neurotransmitter. The direct involvement of GLUD in the pathogenesis of certain human neurodegenerative disorders has been suggested recently. To investigate its possible role in the induction and progression of these disorders, we have initiated studies focusing on the chromosomal organization of the several members of the GLUD family and their functional status. In the present study using a panel of human x rodent somatic cell hybrids and in situ hybridization to metaphase chromosomes, we documented that the members of the GLUD gene family are dispersed in the human genome. The functional GLUD1 gene was mapped to chromosome 10q22.3-q23, and an intronless processed gene (GLUDP1) to chromosome Xq22-q23, while the truncated intron-containing GLUD pseudogene GLUDP2 was also assigned on chromosome 10, but not closely linked to the GLUD1 gene. These results provide novel information concerning the chromosomal organization of the human GLUD gene family.

Animals↗

[A comparative chromosome map between human and Hylobates hoolock built by chromosome painting].

We established chromosomal homologies between all human chromosomes except Y and those of Hylobates hoolock by chromosome painting (chromosome in situ suppression hybridization with human chromosome-specific DNA libraries). Human chromosomes 9,13,15,18,21,22,X libraries hybridized to single G-banding H. hoolock chromosomal region, other human DNA libraries showed at least two homologous segments in H. hoolock chromosomes. 59 segments homologous to one of 22 human autosomes were detected in 18 H. hoolock autosomes. There were at least 39 translocations. Thus an uniquivocally high degree of synteny between human and H. hoolock was confirmed. This will finally allow H. hoolock to be integrated into the overall picture of chromosomal evolution in Hylobatids.

Animals↗

Salivary polytene chromosome mapping of Anopheles (Cellia) subpictus Grassi (Culicidae: Diptera).

With the introduction of molecular taxonomy of mosquitoes, polytene chromosome maps have become indispensable as standard references for locating genes, puffs, and inversion breakpoints of unique DNA sequences. We present a line map and a photomap of the salivary polytene chromosomes of Anopheles (Cellia) subpictus Grassi, an important emerging vector of malaria in India. In addition, we discuss the nature of this species complex consisting of sibling species A, B, C, and D. The comparative study is in relevance to the X chromosome heterozygous inversion differences between 2 allopatric populations of the species and the recognition of 4 X-chromosome inversion genotypes viz: species A-X(+a+b), B-X(ab), C-X(a+b) and D-X(+ab).

Animals↗

Polytene chromosome maps in four species of tsetse flies Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans (Diptera: Glossinidae): a comparative analysis.

Photographic polytene chromosome maps from pupal trichogen cells of four tsetse species, Glossina austeni, G. pallidipes, G. morsitans morsitans and G. m. submorsitans were constructed and compared. The homology of chromosomal elements between the species was achieved by comparing banding patterns. The telomeric and subtelomeric chromosome regions were found to be identical in all species. The pericentromeric regions were found to be similar in the X chromosome and the left arm of L1 chromosome (L1L) but different in L2 chromosome and the right arm of L1 chromosome (L1R). The L2 chromosome differs by a pericentric inversion that is fixed in the three species, G. pallidipes, G. morsitans morsitans and G. m. submorsitans. Moreover, the two morsitans subspecies appeared to be homosequential and differ only by two paracentric inversions on XL and L2L arm. Although a degree of similarity was observed across the homologous chromosomes in the four species, the relative position of specific chromosome regions was different due to chromosome inversions established during their phylogeny. However, there are regions that show no apparent homology between the species, an observation that may be attributed to the considerable intra--chromosomal rearrangements that have occurred following the species divergence. The results of this comparative analysis support the current phylogenetic relationships of the genus Glossina.

Animals↗

Chromosome mapping of genes on the short arm of human chromosome 11: parathyroid hormone gene is at 11p15 together with the genes for insulin, c-Harvey-ras 1, and beta-hemoglobin.

The human parathyroid hormone gene (PTH) was mapped to the 11p15 chromosomal band by in situ hybridization. Using the same procedures and cells, the closely linked beta-hemoglobin gene (HBB), the Harvey-ras 1 proto-oncogene (HRAS1), and the insulin gene (INS) were also mapped to this same region. Some reports have demonstrated differences in regional localization of the latter three genes, and linkage and molecular studies have not resolved how far this linkage group extends from p15 toward the centromere on the physical gene map. Our results show that all of these genes are localized at 11p15, a region of one chromosomal band that appears to comprise a genetic distance of more than 20 cM.

Chromosome Mapping↗

Chromosomal mapping of Adam9, Adam15 and Adam21.

Adam9, Adam15 and Adam21, genes encoding members of the ADAM or MDC family of metalloproteases, have been mapped to mouse chromosomes 8, 1, and 12, respectively, using an interspecific cross. The mapping of these mouse loci and the extrapolated loci for their human orthologs may facilitate the mapping of diseases involving these genes.

ADAM Proteins↗

Chromosome mapping of the human gene encoding the 68-kDa nuclear antigen (p68) by using the polymerase chain reaction.

Chromosome mapping by Southern analysis of DNA from somatic cell hybrids is often unsuccessful when only cDNA probes are available. p68 is a putative RNA helicase that is antigenically related to the simian virus 40 large tumor antigen. By using the polymerase chain reaction and oligode-oxynucleotide primers based on the cDNA sequence, we have identified introns in the p68 gene. Comparison of human and mouse DNA fragments amplified with these primers revealed length differences that allowed us to identify the human gene. Application of this technique to DNA from human-mouse somatic cell hybrids and cell lines derived from them by chromosome-mediated gene transfer allowed us to map p68 to the distal part of the long arm of chromosome 17.

Amino Acid Sequence↗

Structure, exon pattern, and chromosome mapping of the gene for cytosolic copper-zinc superoxide dismutase (sod-1) from Neurospora crassa.

A 4.8-kilobase BamHI-HindIII fragment encoding the entire Neurospora crassa CuZn superoxide dismutase gene (herein designated sod-1) was isolated from a genomic library using two 60-base deoxyoligonucleotide probes corresponding to the published N. crassa amino acid sequence. The nucleotide sequence of the gene encodes an amino acid sequence matching the published protein sequence at 152 of 153 positions. Codon preference shows an unusually strong bias such that only 32 of the possible 61 codons are used, with no codons ending in A. Codon usage is that of highly expressed N. crassa genes. The gene contains three introns, none of which corresponds to any of the introns previously identified in the human gene. Analysis of the intron positions provides support for the hypothesis that CuZn superoxide dismutases evolved by gene duplication and fusion followed by the addition of exons encoding an N-terminal beta-hairpin and a zinc-binding subdomain. The N. crassa gene has an intron mapping to amino acid residue 114 in a sequence-conserved region of the protein whereas the human gene has an intron mapping to a similar but not identical position at residue 118. The discordant position of these introns suggests that one of them was inserted relatively recently. The first N. crassa intron contains a sequence that is similar to the transcriptional regulatory site, UAS1, of the yeast CYC1 (iso-1-cytochrome c) gene and to a putative UAS from the yeast manganese superoxide dismutase gene. A 10-nucleotide portion of this region also matches exactly a sequence in intron 2 of the con-10 gene of N. crassa. sod-1 was mapped to the left arm of chromosome I by following the segregation of a restriction fragment length polymorphism in a sexual cross. Although results indicate that there is a single gene for cytosolic CuZn superoxide dismutase, two additional, perhaps distantly related, sequences were identified that hybridized weakly to both oligonucleotide probes.

Amino Acid Sequence↗