bluuebunny/crossref_metadata_2025 huggingface dataset:
Staining of human telomeres with primed in situ labeling (PRINS)
Authors: Therkelsen A.J. , Nielsen A. , Koch J. , Hindkjær J. , Kølvraa S.
Published: May 2008

As described, the PRINS method is a very rapid and reliable way of staining human telomeres. To obtain the maximum frequency of stained telomeres, the primer (CCCTAA)7 should be used, although the average frequency never quite reaches 100 %. The frequency is strongly dependent on the age of the individual, being significantly higher in children and newborns than in adults. A difference between the (CCCTAA)7 primer and the complementary primer is demonstrated and a possible explanation is proposed, namely, that gaps in the C-rich strand cause chain elongation termination after the addition of only one dTTP molecule.
Localization of the human gene for fibulin-1 (FBLN1) to chromosome band 22q13.3
Authors: Korenberg J.R. , Chen X.-N. , Tran H. , Argraves W.S.
Published: May 2008

Using fibulin-1 cDNA probes, we performed fluorescence in situ hybridization to map the human chromosomal location of the gene encoding the extracellular matrix and blood glycoprotein, fibulin-1 (FBLN1). The gene for fibulin-1 was mapped to a single site on the long arm of human chromosome 22 (22ql3.3). The assignment of the chromosomal map position for the fibulin-1 gene will aid in the evaluation of its potential roles in human connective tissue and blood diseases.
The development of painting probes for dual-color and multiple chromosome analysis in the mouse
Authors: Breneman J.W. , Swiger R.R. , Ramsey M.J. , Minkler J.L. , Eveleth J.G. , Langlois R.A. , Tucker J.D.
Published: May 2008

The recent development of mouse chromosome painting probes for fluorescence in situ hybridization has extended the use of this common laboratory mammal in cytogenetics. We now report the development of additional painting probes by degenerate-oligonucleotide-primed PCR on chromosomes from mouse lung fibroblast cultures, each homozygous for a single Robertsonian translocation chromosome. These probes are for Rb(1.2), Rb(1.3), Rb(4.6), and Rb(6.7). Probes were also made for the sex chromosomes by isolating shoulders from larger peaks (X) or small, clearly resolved peaks (Y) in the flow karyotype. Combinations of probes were used to paint four chromosomes simultaneously in a single color. Multicolor painting was achieved with a biotinylated Rb( 1.2) probe and a digoxigenin-labeled Rb(2.8) probe. Each of the three different homologous pairs was uniquely colored by avidin-Texas Red, anti-digoxigenin-FITC, or both simultaneously. These results extend the usefulness of the mouse as a model for understanding adverse environmental exposures and genetic diseases in humans.
A proposed system for scoring structural aberrations detected by chromosome painting
Authors: Tucker J.D. , Morgan W.F. , Awa A.A. , Bauchinger M. , Blakey D. , Cornforth M.N. , Littlefield L.G. , Natarajan A.T. , Shasserre C.
Published: May 2008

The advent of chromosome painting has brought the realization that structural aberrations can be far more complicated than previously imagined. Various investigators have devised their own nomenclature systems to deal with this difficulty, with the result that the terminology has become inconsistent and confusing. Recently, an international group of cytogeneticists experienced in chromosome painting gathered to address this issue. Results of the meeting are presented in this report, which provides a nomenclature system capable of describing chromosome aberrations that occur between painted and unpainted chromosomes, as well as aberrations involving only painted chromosomes. The nomenclature is flexible enough to describe accurately even the most extensively rearranged chromosomes. As a consequence of this flexibility, the scheme upon which the nomenclature is based differs substantially from other systems of aberration classification. We call this system the Protocol for Aberration Identification and Nomenclature Terminology (PAINT).
The human OTF1 locus which overlaps the CD3Z gene is located at 1q22→q23
Authors: Sturm R.A. , Eyre H.J. , Baker E. , Sutherland G.R.
Published: May 2008

The human OTF1 locus encoding the Oct-1 protein has previously been mapped to chromosome 1cen→q32 by analysis of somatic cell hybrids. We report here the regional localization of OTF1 to 1q22→q23 by fluorescence in situ hybridization. The physical linkage of the CD3Z gene to the OTF1 transcription unit also refines the location of the CD3Z locus to 1q22→q23.
Localization of the undulin gene (UND) to human chromosome band 8q23
Authors: Schnittger S. , Herbst H. , Schuppan D. , Dannenberg C. , Bauer M. , Fonatsch C.
Published: May 2008

Undulin, a large extracellular matrix (ECM) mosaic glycoprotein related to collagen type XIV, is associated with dense collagen matrices in soft tissues and is likely to be involved in the supramolecular organization of interstitial collagens. By fluorescence in situ hybridization (FISH), we have assigned the undulin locus (UND) to band q23 of human chromosome 8.
Localization of the human HTF4 transcription factors 4 gene (TCF12) to chromosome 15q21
Authors: Zhang Y. , Flejter W.L. , Barcroft C.L. , Rivière M. , Szpirer J. , Szpirer C. , Bina M.
Published: May 2008

Identification and localization of genes that encode regulators of transcription could provide landmarks for functional analysis of the human genome. Toward this goal, we examined a panel of somatic cell hybrids and assigned the gene (TCF12) encoding the helix-loop-helix transcription factors 4 (HTF4) to chromosome 15. Fluorescence in situ hybridization further localized TCF12 to chromosome 15q21. Northern analysis revealed that the relative abundance of HTF4 gene transcripts is not constant but varies depending on the human cell-line or tissue examined.
Fine deletion mapping of the p22 region of the human X chromosome using a radiation-induced hybrid panel
Authors: Heuertz S. , Smahi A. , Sanak M. , Holvoet-Vermaut L. , Hors-Cayla M.-C.
Published: May 2008

Radiation-induced somatic cell hybrids containing fragments of the human X chromosome were constructed. A panel of 17 hybrids was selected with the help of known markers in the Xp22 region. These hybrids identified 11 different breakpoints between Xp22.2 and Xp21.3. Eight markers were located in eight of the nine corresponding intervals, resulting in the following physical map: tel DXS89-DXS278-DXS85-(DXS1224, DXS16)-(GLRA2, DXS987)-DXS207-(DXS-197, DXS1053)-(DXS43, DXS1195)-(DXS1229, DXS-999)-(DXS1052, DXS92, DXS274)-(DXS41, DXS1226)-DXS1198-DXS28...cen.
Chromosome banding in Amphibia
Authors: Schmid M. , Feichtinger W. , Weimer R. , Mais C. , Bolaños F. , León P.
Published: May 2008

Cytogenetic analyses were performed on several populations of the Central American tree frog Agalychnis callidryas, using conventional methods and banding techniques. The karyotype of this species is distinguished by an inversion polymorphism in chromosome 9, which is either submetacentric or telocentric. The populations examined are in Hardy-Weinberg equilibrium with respect to the two alternative morphs of chromosome 9. This is the first report of the occurrence of an intrapopulational chromosomal inversion polymorphism in the order Anura. In male meiosis, the two chromosomes 9 form a bivalent exhibiting a ring-like pairing configuration with terminal chiasmata in both arms, regardless of whether the paired homologs are heteromorphic or homomorphic. Furthermore, individual specimens of A. callidryas exhibit one or two unexpected 18S + 28S ribosomal RNA gene clusters, in addition to the standard nucleolus organizers. The chromosomal localization of these extra nucleolus organizers is identical in all metaphases from the same specimen and shows a specific intraindividual pattern. The karyotype evolution in the phyllomedusine hylids, the structure of the various classes of heterochromatin, and the occurrence and possible origin of the rare inversion polymorphisms and multiple nucleolus organizers in A. callidryas and a few other amphibian species are discussed.
Mapping of an ordered set of 14 cosmids to human chromosome 12p by two-color in situ hybridization
Authors: Chaffanet M. , Baens M. , Aerssens A. , Schoenmakers E. , Cassiman J.-J. , Marynen P.
Published: May 2008

To map human chromosome 12p aberrations by fluorescence in situ hybridization (FISH), cosmids were isolated or obtained for 14 known 12p loci (D12S119, D12S158, D12S178, D12S370, D12S380E, A2M, CACNLlAl, FGF6, GAPD, KRAS2, PRB1, PZP, TPI1, and VWF). Using two-color FISH with three labeled probes to interphase nuclei, and to prometaphase chromosomes where possible, the order of these loci was sequentially determined to be pter-D12S158- D12S380E-CACNL1A1-FGF6-D12S370-VWF-GAPD-TPIl-A2M-PZP-PRBl-D12S178-D12S119-KRAS2-cen. Two cell lines were analyzed with this set of cosmids. The EBV-transformed cell line TA carries a der(12) with a deletion of bands 12p13.1→p11.2. D12S178, D12S119, and KRAS2 were absent in the der(12), whereas the other loci were present. The second cell line, GM01203A, exhibits a balanced t(4;12)(4q25; 12p13.3) with a breakpoint between FGF6 and D12S370.
Chromosomal localization of the BLV receptor candidate gene in cattle, sheep, and goat
Authors: Popescu C.P. , Boscher J. , Hayes H.C. , Ban J. , Kettmann R.
Published: May 2008

The BLV receptor candidate gene has been localized precisely on cattle, goat, and sheep chromosomes using nonradioactive in situ hybridization and simultaneous fluorescent R-banding. The probe, a whole plasmid containing a 2.3-kb bovine cDNA fragment encoding part of this gene, was nick-translated in the presence of biotin-11-dUTP. It hybridized on band q15 of cattle chromosome 7, goat chromosome 7, and sheep chromosome 5, thus confirming the homoeology based on banding patterns among the chromosomes of these three species.
Cloning and regional assignment of the human myosin heavy chain 12 (MYH12) gene to chromosome band 15q21
Authors: Moore K.J. , Testa J.R. , Francke U. , Milatovich A. , Copeland N.G. , Jenkins N.A.
Published: May 2008

Sequences encoding 1,235 bp of the human myosin heavy chain 12 (MYH12) gene have been cloned from a human brain cDNA library by PCR amplification. The human sequence is 95.8% identical to the mouse sequence at the amino acid level, indicating that the MYH12 gene has been evolutionarily well conserved. Somatic cell hybrid analysis and in situ hybridization place the MYH12 gene on human chromosome 15, at band q21, and extend distally the known region of chromosome 15 linkage homology on mouse chromosome 9.
Absence of terminal telomeric FISH signals in chromosomes from immortal Chinese hamster cells
Authors: Slijepcevic P. , Bryant P.E.
Published: May 2008

In contrast to normal Chinese hamster cells, terminal telomeric signals were not detectable by fluorescence in situ hybridization (FISH) in their immortalized counterparts, even when a sensitive cooled CCD camera was used for acquiring FISH images. Since the telomeres were detectable by FISH in immortal human cells, this suggests that hamster and human cells might have different mechanisms of telomere processing.
Assignment of Tp53 and Tk1 to chromosome 13 in Peromyscus by fluorescence in situ hybridization
Authors: Wang Z. , Young S.R. , Liu L. , Dawson W.D.
Published: May 2008

Mus domesticus DNA probes for the tumor suppressor protein-5 3 (Tp53) and thymidine kinase-1 (Tkl) genetic loci were used to identify clones representing these loci in a Peromyscus leucopus (white-footed mouse) cosmid library. The cosmid-derived homologous probes were biotinylated and hybridized to P. mαniculatus (deer mouse) chromosomes. Probes for both genes hybridized to the chromosome 13 pair identified by prior G-banding. Deer mouse chromosome 13 shares a region of homology with mouse chromosome 11, rat chromosome 10 and human chromosome 17.
Description of a 700-kb yeast artificial chromosome contig containing the BCL1 translocation breakpoint region at 11q13
Authors: Szepetowski P. , Perucca-Lostanlen D. , Grosgeorge J. , LePaslier D. , Brownstein B.H. , Carle G.F. , Gaudray P.
Published: May 2008

We screened two human yeast artificial chromosome (YAC) libraries by polymerase chain reaction (PCR) with oligonucleotides specific to the BCL1 major translocation breakpoint cluster region at 11q13. Five YACs were isolated. Two of them were chimeric. One of these and remaining three YACs were characterized by hybridization with various known 11q13 probes, Alu-PCR fingerprinting, in situ hybridization, and isolation of YAC ends. A map of this ca 700-kb YAC contig was obtained. This map was consistent with maps established from total human genomic DNA. Every YAC in this region was found unstable and gave rise to reproducibly deleted lineages. Analysis in detail of these deletions over many generations showed that more than a single sequence might be involved. The availability of cloned material will facilitate the search for the still elusive genetic elements responsible for amplifications, deletions and translocations observed at 11q13 in malignancies.
The gene (LGALS3BP) encoding the serum protein 90K, associated with cancer and infection by the human immunodeficiency virus, maps at 17q25
Authors: Calabrese G. , Sures I. , Pompetti F. , Natoli G. , Palka G. , lacobelli S.
Published: May 2008

Levels of a 90-kDa tumor associated protein, designated 90K (gene symbol LGALS3BP), have been found elevated in the serum of patients with cancer and in those infected by the human immunodeficiency virus (HIV). 90K appears to be implicated in immune response associated with natural killer (NK) and lymphokine-activated killer (LAK) cell cytotoxicity. Using fluorescence in situ hybridization the full length 90K cDNA has been localized to chromosome 17q25.
Mouse chromosome 6 in Rb translocations: consequences in singly and doubly heterozygous males
Authors: Aranha I.P. , Martin-DeLeon P.A.
Published: May 2008

Previously it has been shown that male mice heterozygous for the Rb(6.16) translocation show a transmission distortion of the segregants in resulting zygotes, particularly for sexually active animals (Aranha and Martin-DeLeon, 1991). Sperm bearing the translocation have an impaired fertilizing ability, compared to chromosomally normal sperm, and this dysfunction is potentiated in X-bearing cells leading to a sex ratio distortion. Since earlier data suggested that a deficiency of Chromosome 6 was associated with sperm dysfunction (Zackowski and Martin-DeLeon, 1989), the present study was designed to determine the uniqueness of the coupling of the distortions for the Rb(6.16) translocation and the effect of 0 and 2 copies of Chr 6 on sperm function. First cleavage metaphases, resulting from chromosomally normal females that were mated to sexually active or rested Rb(6.15) or Rb(2.8) heterozygous males, were sequentially G-and C-banded. For unaged sperm from sexually active Rb(6.15) males, cytogenetic analysis showed that the ratio of normal (90) to balanced (37) zygotes deviated significantly (P < 0.001) from 1:1, as did the sex ratio, 46X:81Y (P < 0.01). The latter distortion was related to a deficiency of X-bearing sperm with the translocation (8X:29Y). Both segregation and sex ratio distortions disappeared for aged sperm (69:50, 51X:68Y) and a contingency table showed that segregant type was related to sperm age (P < 0.05). For the Rb(2.8) the ratio of normal to balanced zygotes (65:46) was not significantly different from 1:1 for unaged sperm, and although the ratio from aged sperm was 62:39, there was no relationship between segregant type and sperm age (P > 0.05). There was also no sex ratio distortion for either unaged or aged sperm (P > 0.05). To generate zygotes with sperm complements containing 0 and 2 copies of Chr 6, males doubly heterozygous for Rb(6.16) and Rb(6.15) were mated to chromosomally normal females. No zygotes were recovered and the males were shown histologically to have azoospermia and spermatogenic arrest. The results support the conclusion that the presence of Chr 6 in Rb translocations specifically affects segregant and sex ratio distributions. They also indicate, that as the common arm in compound heterozygotes, this chromosome grossly affects spermatogenesis.
Chromosome banding patterns in the unisexual microteiid Gymnophthalmus underwoodi and in two related sibling species (Gymnophthalmidae, Sauria)
Authors: Yonenaga-Yassuda Y. , Vanzolini P.E. , Rodrigues M.T. , de Carvalho C.M.
Published: May 2008

Cytogenetic investigation of three species of the genus Gymnophthalmus (the unisexual G. underwoodi, G. leucomystax and Gymnophthalmus sp.n., a new, undescribed redtailed species) documented three different karyotypes, all with 2n = 44. Chromosomes from fibroblast cultures were studied after routine Giemsa staining, CBG-banding, RBG-banding, and Ag-NOR staining. The unisexual species showed a very distinctive karyotype from the one previously described that, paradoxically, corresponded to the predicted karyotype of one of its parental species. Our chromosome data unequivocally show that there are two parthenogenetic species with two different mechanisms of origin under the name underwoodi.
Human origin of micronuclei in human × hamster two-cell embryos
Authors: Tusell L. , Alvarez R. , Genescà A. , Caballín M.R. , Miró R. , Barrios L. , Egozcue J.
Published: May 2008

Using fluorescence in situ hybridization techniques with either human or hamster genomic DNA probes, we studied the origin of micronuclei in two-cell hybrid embryos obtained from hamster oocytes and γ-irradiated human spermatozoa. Our study demonstrates that over 99 % of micro nuclei hybridize with human DNA probes and not with hamster DNA, revealing their human origin. Thus, the micronucleus test represents a good method to evaluate genetic damage in human germ cells, since it is simpler and faster than sperm chromosome studies.
Dehydroepiandrosterone sulfotransferase gene (STD): localization to human chromosome band 19q13.3
Authors: Otterness D.M. , Mohrenweiser H.W. , Brandriff B.F. , Weinshilboum R.M.
Published: May 2008

Dehydroepiandrosterone (DHEA) sulfotransferase (ST) catalyzes the sulfate conjugation of DHEA and other steroid compounds. The human gene for DHEA ST (STD) was mapped by the polymerase chain reaction to chromosome 19 using human × rodent somatic cell hybrid panels. Fluorescence in situ hybridization was then used to localize the STD gene to the region 19q13.3.
Assignment of the human protein tyrosine phosphatase, receptor-type, zeta (PTPRZ) gene to chromosome band 7q31.3
Authors: Ariyama T. , Hasegawa K. , Inazawa J. , Mizuno K. , Ogimoto M. , Katagiri T. , Yakura H.
Published: May 2008

Human protein tyrosine phosphatase, receptortype, zeta (PTPRZ; also denoted HPTPζ or RPTPβ) has a large extracellular region with the N-terminal carbonic anhydrase-like domain and a cytoplasmic region with two tandemly located protein tyrosine phosphatase domains. One of the characteristics of PTPRZ is its preferential expression in the central nervous system. We localized the human PTPRZ gene to chromosome band 7q31.3 by somatic cell hybrid mapping and fluorescence in situ hybridization.
Assignment of the human β-catenin gene (CTNNB1) to 3p22→p21.3 by fluorescence in situ hybridization
Authors: van Hengel J. , Nollet F. , Berx G. , van Roy N. , Speleman F. , van Roy F.
Published: May 2008

The human β-catenin gene (CTNNB1) has been localized to 3p22→p21.3 by fluorescence in situ hybridization. Recent studies have suggested the presence of one or more tumor suppressor genes on the short arm of chromosome 3. This raises the possibility that CTNNB1, for which important features are already known, is involved in tumor progression.
Additional assignment of the human 5S rRNA genes to chromosome region 1q31
Authors: Lomholt B. , Frederiksen S. , Nederby Nielsen J. , Hallenberg C.
Published: May 2008

A major fraction of the human 5S rRNA genes has previously been assigned to chromosome region 1q42.11→q42.13 (Sørensen et al., 1991). Through in situ hybridization of different tritiated probes to metaphase spreads, we have demonstrated that a minor fraction of the 5S rRNA genes is localized at band lq31. This fraction amounts to 25–30% of the genes found in the region 1q42.11→q42.13. Results obtained with the chromosomes of a balanced translocation carrier involving this region indicate that the major cluster is localized in band 1q42.13.
Segregation analysis of four translocations, t(2;18), t(3;15), t(5;7), and t(10;12), by sperm chromosome studies and a review of the literature
Authors: Estop A.M. , Van Kirk V. , Cieply K.
Published: May 2008

We examined the meiotic segregation patterns of 444 sperm cells belonging to four reciprocal translocation carriers, t(2;18)(p21;q11.2), t(3;15)(q26.2;q26.1), t(5;7)(q13;p15.1), and t(10;12)(q26.1;p13.3). For the t(2;18) carrier, the frequencies of alternate, adjacent-1, adjacent-2, and 3:1 segregations were 41.9%, 35.2%, 14.4%, and 8.4%, respectively. For the t(3;15) carrier, the segregation pattern was 48% alternate, 36% adjacent-1, 12% adjacent-2, 2% 3:1, and 2% 4:0. One cell was the result of a 4:0 segregation. For the t(5;7) heterozygote, the corresponding segregation frequencies were 40.2%, 26.2%, 16.6%, and 17.0%. This translocation heterozygote showed a higher number of 3:1 segregations than adjacent-2 segregations, which is unusual. The t(10;12) segregations were 61.1%, 26.3%, 6.9%, and 5.6%. The percentages of chromosome abnormalities unrelated to the translocation ranged from 0% to 0.6% for aneuploidy and from 5.5% to 10.9% for structural abnormalities. These frequencies are within the ranges for control donors. Sperm chromosome data from the literature on the segregation of 30 reciprocal translocations were reviewed.
A microdissection library of the rat renal carcinoma gene region
Authors: Bell D.W. , Yeung R.S. , Bohlander S.K. , Cheng J.Q. , Jin F. , Testa J.R.
Published: May 2008

Predisposition to hereditary renal carcinoma in the Eker rat involves a mutation of a putative tumor suppressor gene within chromosome band 10q12. We describe the identification of three unique polymorphic sequences in the vicinity of this locus following the microdissection, construction and characterization of a region-specific DNA library for rat chromosome band 10ql2.
Construction of Chinese hamster chromosome specific DNA libraries and their use in the analysis of spontaneous chromosome rearrangements in different cell lines
Authors: Balajee A.S. , Domínguez I. , Natarajan A.T.
Published: May 2008

Chinese hamster (Cricetulus griseus) and a wide variety of cell lines derived from it have been extensively used for radiobiological and genotoxicity studies. In this report, we describe the development of chromosome-specific DNA libraries for Chinese hamster chromosomes 1, 2, 3, 5, 6, 7 and 8 by the linker/adaptor method. The specificity of these probes have been checked by FISH on primary embryonic fibroblast (CHE) metaphase chromosomes. The diploid number of chromosomes in different cell lines of CHO often varies from 19 to 21 and karyotyping is very difficult owing to many spontaneous rearrangements. Using Chinese hamster chromosome-specific DNA libraries, we have analysed the spontaneous chromosome rearrangements in three different Chinese hamster transformed cell lines, V79, CHO-KI and CHO-9. The results indicate that one of each of the first and fifth pair of chromosomes are involved in reciprocal translocations in both CHO-9 and CHO-KI cells. The hybridisation patterns obtained with chromosome-specific probes 2, 3, 5, 6, 7 and 8 indicated rearrangements have occured involving terminal and interstitial translocations in V79, CHO-KI and CHO-9 cells.
Regional localisation of tri- and tetranucleotide repeat sequence-containing cosmids on chromosome 13
Authors: Hawthorn L.A. , Kapanadse B. , Yankovsky N. , Cowell J.K.
Published: May 2008

A human chromosome 13-specific cosmid library has been screened with oligonucleotides containing tri- and tetranucleotide repeats with motifs: GACA, GACT, CAC, TCC. 20 cosmid clones were identified and their physical location on chromosome 13 was determined using fluorescence in situ hybridisation. Over 80% of the cosmids were detected using tetranucleotide repeats of which 50% were GACT, but hese were distributed along the length of the chromosome. Clones from the 13q12→q14 and 13q32→q34 regions were apparently overrepresented and 10% of cosmids were localised to the short arm of the chromosome. These simple tandem repeats will be an important addition to the genetic mapping strategy for human chromosome 13.
Molecular cytogenetic analysis of the double-CMA3 chromosome of lake trout, Salvelinus namaycush
Authors: Reed K.M. , Phillips R.B.
Published: May 2008

The chromosome possessing two chromomycin A3 (CMA3) staining sites in lake trout (Salvelinus namaycush) was examined using FISH. All CMA3-bands in the karyotype contained ribosomal DNA (rDNA), including both sites on the double-CMA3 chromosome. One rDNA site on this chromosome was bordered by telomeric repeats, implicating a rearrangement in the origin of the second NOR.
Regional localization of the ovine NRAMP gene to chromosome 2q41→q42 by in situ hybridization
Authors: Pitel F. , Cribiu E.P. , Yerle M. , Lahbib-Mansais Y. , Lanneluc I. , Lantier F. , Gellin J.
Published: May 2008

The murine Ity/Lsh/Bcg locus belongs to a conserved region between mouse Chromosome 1 and human chromosome 2. This gene governs mouse resistance to several intracellular pathogens. We have localized the gene on sheep chromosome 2q41→q42 by radioactive in situ hybridization using a 2.1-kb fragment of the ovine equivalent of this gene.
Determination of the gene order of the three loci CD2, NGFB, and NRAS at human chromosome band 1p13 and refinement of their localisation at the subband level by fluorescence in situ hybridisation
Authors: Mitchell E.L.D. , Jones D. , White G.R.M. , Varley J.M. , Santibanez Koref M.F.
Published: May 2008

The three loci NRAS, NGFB, and CD2 map toband 1p13.2 and CD2 and NGFB to human chromosome band 1p13. Using fluorescence in situ hybridisation (FISH) to simultaneously DAPI-banded metaphase chromosomes, we have further refined the localisation of these three genes to specific subbands. NRAS localises to sub-1p13.1. Also, with the use of multicolour FISH, we have determined the order and orientation of the three loci in relation to the centromere. The orders cen-CD2-NGFB-NRAS.
Assignment of α1-acid glycoprotein gene (ORM1 )to swine chromosome region 1q210→q212 by fluorescence in situ hybridization
Authors: Murakami Y. , Itoh T. , Ohata K. , Yasue H.
Published: May 2008

The porcine gene for α1-acid glycoprotein (ORM1) was localized to chromosome region 1q210→ q212 by fluorescence in situ hybridization. From this result and published gene mapping data, it was concluded that the syntenic group comprised of ORM1, IFNA1, and GRP78 in the human genome is also conserved in the pig.
The human gonadotropin-releasing hormone receptor gene (GNRHR) maps to chromosome band 4q13
Authors: Kakar S.S. , Neill J.D.
Published: May 2008

A cDNA representing the high-affinity gonadotropin-releasing hormone (GnRH) receptor has been molecularly cloned from the human pituitary gland, a breast tumor cell line (MCF 7), and an ovarian tumor. The nucleotide sequence of this cDNA was determined, and its expression in various human tumors and tumor cell lines was demonstrated. In this study, we localized the gene encoding the GnRH receptor to human chromosome 4, using polymerase chain reaction (PCR) analysis of genomic DNA from human × hamster somatic cell hybrids. The gene was sublocalized to chromosome band 4q13 using fluorescence in situ hybridization with the GnRH receptor gene (GNRHR).
Localization of the human AHNAK/desmoyokin gene (AHNAK) to chromosome band 11q12 by somatic cell hybrid analysis and fluorescence in situ hybridization
Authors: Kudoh J. , Wang Y. , Minoshima S. , Hashimoto T. , Amagai M. , Nishikawa T. , Shtivelman E. , Bishop J.M. , Shimizu N.
Published: May 2008

Using a panel of somatic cell hybrids, we have mapped the locus for the AHNAK/desmoyokin gene (AHNAK) to human chromosome 11. Fluorescence in situ hybridization experiments independently confirmed the chromosomal localization and refined it to band 1 1q12.
Chromosomal assignment of the human deoxyribonuclease I gene, DNASE1 (DNL1), to band 16p13.3 using the polymerase chain reaction
Authors: Yasuda T. , Nadano D. , Iida R. , Takeshita H. , Lane S.A. , Callen D.F. , Kishi K.
Published: May 2008

To localize the human deoxyribonuclease I (DNase I) gene, DNASE1 (DNL1), we performed a polymerase chain reaction (PCR) using DNA extracted from a panel of cloned human × rodent hybrid cell lines carrying different human chromosomes and screened for the presence of the expected PCR products. Two different sets of oligonucleotide primers specific for human DNase I cDNA sequences were used to amplify unique fragments in the human DNase I gene. Based on this work, DNL1 could be assigned to human chromosome 16. Furthermore, regional localization of the gene to 16p13.3 was performed by PCR analysis of a high-resolution mouse × human somatic cell hybrid panel that contained defined portions of human chromosome 16.
The human calcitonin receptor gene (CALCR) at 7q21.3 is outside the deletion associated with the Williams syndrome
Authors: Pérez Jurado L.A. , Li X. , Francke U.
Published: May 2008

The human calcitonin receptor (CTR) is a transmembrane peptide with dual action as a receptor for the hormone calcitonin and as an extracellular calcium sensor. Therefore, CTR dysfunction could lead to disorders of calcium metabolism associated with hypercalcemia, such as the Williams syndrome (WS). WS is a developmental disorder caused by a deletion at chromosome 7q11.23 that includes the elastin locus (ELN). We have mapped the CTR gene (CALCR) to chromosome band 7q21.3 by polymerase chain reaction and singlestrand conformation analysis of somatic cell hybrids as well as fluorescence in situ hybridization (FISH) to metaphase chromosome spreads. Two-color FISH cohybridizing CTR and ELN probes confirmed that CALCR maps telomeric to ELN. Subsequent analysis of chromosome spreads from four WS patients revealed deletion of the ELN locus in all of them and normal hybridization of CTR probes to both chromosome 7 homologues, indicating that CALCR lies outside the deleted region.
Chromosomal mapping of the genes GPRK5 and GPRK6 encoding G protein-coupled receptor kinases GRK5 and GRK6
Authors: Bullrich F. , Druck T. , Kunapuli P. , Gomez J. , Gripp K.W. , Schlegelberger B. , Lasota J. , Aronson M. , Cannizzaro L.A. , Huebner K. , Benovic J.L.
Published: May 2008

G protein-coupled receptor kinases (GRKs) play an important role in phosphorylating and regulating the activity of a variety of G protein-coupled receptors. Chromosomal mapping of the human genes for the two most recently identified members of the GRK family, GRK5 (GPRK5) and GRK6 (GPRK6), was accomplished by correlation of the presence of the GPRK5 and GPRK6 loci with specific chromosome regions in a rodent-human hybrid panel. These analyses revealed that GPRK5 maps to chromosome region 10q24→qter while GPRK6 maps to 5q35. A GPRK6 related locus maps to 13pter→q21.
Rabl orientation of CENP-B box sequences in Tupaia belangeri fibroblasts
Authors: Haaf T. , Ward D.C.
Published: May 2008

The chromosomes of the tree shrew Tupaia belangeri exhibit highly localized CENP-B box sequences in the centromeric regions of most chromosomes. Telomeric sequences are present at the ends of all chromosomes and, in addition, at specific interstitial chromosomal sites that likely represent remnants of ancestral telomeres. This suggests that Robertsonian and tandem chromosome fusion events have occurred in the karyotypic evolution of Tupaiidae. In Tupαiα skin fibroblasts CENP-B boxes are almost always clustered together at one pole of the interphase nucleus, whereas the telomeric domains are relatively evenly distributed throughout the whole nuclear volume. The observed orientation of the centromeres is reminiscent of the Rabl polarization of chromosomes; this is the first mammalian cell substrate in which such an higher-order chromosomal organization has been observed. CENP-B box sequences are found in several other mammalian species. The implications for recent parallel evolution of CENP-B binding motifs and concerted evolution of these sequences are discussed.
Assignment of the gene (GLCLC) that encodes the heavy subunit of γ-glutamylcysteine synthetase to human chromosome 6
Authors: Sierra-Rivera E. , Summar M.L. , Dasouki M. , Krishnamani M.R.S. , Phillips J.A. , Freeman M.L.
Published: May 2008

Assignment of the human gene (GLCLC) that encodes the heavy or catalytic subunit of γ-glutamylcysteine synthetase (glutamatecysteine ligase; EC 6.3.2.2) to humanchromosome 6 was accomplished by hybridization to Southern blotted somatic cell hybrid DNA. This assignment was confirmed by PCR from somatic cell hybrid DNAs.
Molecular characterization and refined genomic localization of three human potassium ion channel genes
Authors: Landes G.M. , Curran M.E. , Keating M.T.
Published: May 2008

Potassium ion (K+) channels are essential for a variety of cellular functions in both excitable and non-excitable cells and are likely to be involved in the pathogenesis of some cardiovascular and neurological disorders. To be useful in candidate gene analysis of inherited diseases it is important to identify new K+ channel genes and localize these sequences on the human physical and genetic maps. Using fluorescence in situ hybridization (FISH), we mapped two new K+ channel gene containing cosmids, c2-3a and c9-2a, to chromosomes 1 and 19, respectively. Partial DNA sequencing (c2-3a) and restriction enzyme site analysis (c9-2a) established the uniqueness of each clone. We refined the localization of c2-3a, c9-2a and a previously described K+ channel gene KCNA5, (c7-2), by performing contour length measurements of hybridized metaphase chromosomes and determining the average FLpter % value (fractional length relative to the fixed reference point pter × 100%). When compared to ideograms of banded metaphase chromosomes, these FLpter% values correspond to 12p13.31→p13.33, 1p13.1→p21.1 and 19q13.32→q13.33, respectively. Using FISH, each of these clones has been finely mapped to a different human chromosome indicating a significant dispersion of K+ channel sequences in the human genome.
Improved simple generation of GTG-band specific painting probes
Authors: Yokoyama Y. , Sakuragawa N.
Published: May 2008

We developed an improved, simple method of generating chromosome-region-specific probes from only a few microdissected chromosomes. One to five dissected fragments from a defined chromosomal region were processed with a PEG/proteinase K cycling deproteinization step and directly amplified with a two-step amplification system using a degenerate oligonucleotide primed shuttle polymerase chain reaction (DOP-Shuttle-PCR). This modified method offered three advantages over previously reported methods: relaxation of the highly condensed chromosomal DNA, reduction of the risk of endogenous and exogenous contamination, and high efficiency amplification of template DNA. High intensity in the fluorescence in situ hybridization (FISH) signals from normal metaphase chromosomes, as well as regional specificity of these probes, corresponding to regions on R-banded chromosomes, were observed.
Simultaneous visualization of Q-bands and FISH signals using a novel fluorochrome
Authors: Kondoh Y. , Ono T. , Kagiyama N. , Fujita S. , Momiyama M. , Hori S.H. , Yoshida M.C.
Published: May 2008

We developed a new HNPP-azo dye method for detection of fluorescence in situ hybridization (FISH) signals on Q-banded chromosomes by use of a newly synthesized fluorochrome, HNPP (3-hydroxy-N-2’-biphenyl-2-naphthalenecarboxamide phosphate ester), which reacts enzymatically with alkaline phosphatase and azo dye. The FISH staining method permits simultaneous detection of orange HNPP signals on chromosomal sites labeled by Q-banding, allowing the assignment of small (440–1,200 bp) probes.
Selection of chromosome-specific primers and their use in simple and double PRINS techniques for rapid in situ identification of human chromosomes
Authors: Pellestor F. , Girardet A. , Lefort G. , Andréo B. , Charlieu J.P.
Published: May 2008

The PRimed IN Situ labeling (PRINS) technique is an alternative to in situ hybridization for rapid chromosome screening. We have defined and tested new specific oligonucleotide primers for α-satellite DNA of several chromosomes. When using a semiautomatic PRINS protocol, specific labeling was obtained in both metaphase cells and interphase nuclei in a 1-h reaction. PRINS may be a simple and reliable technique for rapidly detecting aneuploidies.
FISH mapping of 22 novel X chromosome cosmids and the isolation of a novel microsatellite on proximal Xp
Authors: Kamakari S. , Thiselton D. , Lindsay S. , Hardcastle A. , Bhattacharya S.
Published: May 2008

Twenty-two X-linked cosmid clones have been localised by fluorescence in situ hybridisation (FISH). Twelve map to the long arm of the X chromosome and 10 to the short arm. Seven of the latter cosmids and an additional one were mapped by two colour FISH relative to reference markers DXS7 and DXS426 in proximal Xp. Finer localisation of one of the cosmids, namely HX43 was achieved by isolation of a microsatellite followed by genetic mapping with respect to reference markers in the region.
Physical linkage of the cdc2-related gene (PCTK1) and the ubiquitin-activating enzyme E1 gene (UBE1) on human Xp11.3
Authors: Knight J.C. , Renwick P.J. , Downing J.R. , Okuda T.
Published: May 2008

PCTAIRE-1 is a cdc2-related protein kinase of unknown function. The gene (PCTK1) has been mapped to chromosome Xp. In this study we refine the locus position by chromosome analysis with cosmid and YAC probes. PCTK1 maps distal to the t(X;18) synovial sarcoma breakpoint in Xp11.23. A 420-kb YAC clone positive for PCTK1 also contains the gene coding for ubiquitin-activating enzyme UBEl, previously mapped in Xp11.3, indicating close physical linkage of these genes. PCTK1 is a new candidate for heritable disorders mapped to the Xp11.3→p11.23 region.
Physical evidence for the position of the Friedreich’s ataxia locus FRDA proximal to D9S5
Authors: Hillermann R. , See C.G. , Pook M. , Wilkes D. , Carvajal J. , Doudney K. , Williamson R. , Chamberlain S.
Published: May 2008

Orientation of the Friedreich’s ataxia locus (FRDA) with respect to D9S15 and D9S5 has proved critical to the design of subsequent cloning strategies. The rarity of recombination events between FRDA and these markers, originally used to determine assignment to human chromosome region 9q13→q21.1 has necessitated the instigation of physical mapping studies to determine order and, hence, the precise location of the disease gene. Simultaneous fluorescence in situ hybridisation using cosmid clones located in close proximity to the ends of a 1.2-Mb yeast artificial chromosome clone extending into the FRDA candidate region provides physical evidence for the order of the marker loci to be cen-D9S202-D9S5-D9S15-qter. The possibility that a pericentric inversion, occurring naturally in ∼1 % of the normal population, may affect the order of markers within this region has been eliminated. Considered in association with the interpretation of a recombination event detected in a single affected individual, these data indicate that the FRDA locus is located proximal to D9S5.
Multicolor FISH with a telomere repeat and Sry sequences shows that Sxr (Sex reversal) in the mouse is a new type of chromosome rearrangement
Authors: Ashley T. , Lieman J. , Ward D.C.
Published: May 2008

XYsxr (sex reversal) mice carry a Y chromosome in which the chromatin (including Sry, the gene for testis determination) that normally resides on the short arm is duplicated and the second copy is relocated to the distal end of the long arm. Multicolor in situ hybridization to mitotic chromosomes of XYsxr males using probes for the telomere repeat sequence (TTAGGG)n and Sry shows that the rearranged chromatin is located distal to the telomeric signal. This suggests that the rearrangement arose from a recombination event involving the distal Y telomere sequences, i.e., within the telomere, a structure historically assumed to be incapable of participating in chromosome rearrangements.
Localization of IGF1R and EDN genes to pig chromosomes 1 and 7 by in situ hybridization
Authors: Lahbib-Mansais Y. , Yerle M. , Gellin J.
Published: May 2008

The genes coding for the insulin-like growth factor-1 receptor (IGF1R) and a porcine endothelium-derived 21-residue vasoconstrictor peptide (EDN) were localized in the pig by means of radioactive in situ hybridization. IGF1R was mapped to chromosome region 1q1.7→q2.1 and EDN to 7p1.3→p1.2. The results are discussed in relation to the position of their homologous loci in man.
NOR and heterochromatin analysis in two cartilaginous fishes by C-, Ag- and RE (restriction endonuclease)-banding
Authors: Stingo V. , Rocco L. , Odierna G. , Bellitti M.
Published: May 2008

Selachian chromosomes were investigated for the first time by C-, Ag- and RE (restriction endonuclease)-band-ing. A pair of NOR-bearing acrocentric chromosomes, as well as a third extra site, were demonstrated in two species of Torpedo. In situ digestion with restriction endonucleases revealed a peculiar arrangement of constitutive heterochromatin: in Torpedo ocellata the centromere never appeared C-banded, and was always digested by AluI, PstI, DdeI and HaeIII; in Torpedo marmorata, instead, heterochromatin was present in the centromere, and was not digested by the same enzymes. The occurrence of different types of centromeric heterochromatin suggests an involvement of highly repeated DNA fractions in the Robertsonian phenomena that occurred in chromosome rearrangements in the genus Torpedo.
Genetic and radiation-reduced somatic cell hybrid sublocalization of the human GSTP1 gene
Authors: Smith C.M. , Bora P.S. , Bora N.S. , Jones C. , Gerhard D.S.
Published: May 2008

A number of related enzymes like glutathione S-transferases (GSTs) and fatty acid ethyl ester synthases (FAEESs) have been implicated in detoxification and drug resistance. The anionic class of GSTs, π, and closely related FAEES-III exhibit tissue-specific and developmentally regulated expression, and the former has been shown to be overex-pressed or amplified in a variety of tumors. The GSTPl gene has previously been cloned and cytogenetically localized to human 11q13 by in situ hybridization. Using a series of previously described radiation-reduced somatic cell hybrids, we have sublocahzed GSTP1 to 11q13. We isolated a genomic clone containing the entire GSTPl gene and sequenced it. Analysis of the 5’ region revealed 23 (TAAAA) tandem repeats interrupted by a single TA and TAA insertion. This repeat number differs among individuals. Eleven alleles in a mostly Caucasian sample were observed. This repeat has a polymorphism information content of 0.74. Linkage analysis of the Venezuelan reference pedigree places GSTP1 5 cM distal to PYGM and 4 cM proximal to FGF3 thereby providing a genetic marker half-way between these two loci. The sublocalization and genetic characterization of GSTPl facilitates linkage analysis of several disease genes mapped to this chromosome band as well as the correlation of genetic and physical markers in the region.
Meiotic segregation in males heterozygote for reciprocal translocations: analysis of sperm nuclei by two and three colour fluorescence in situ hybridization
Authors: Rousseaux S. , Chevret E. , Monteil M. , Cozzi J. , Pelletier R. , Devillard F. , Lespinasse J. , Sèle B.
Published: May 2008

The meiotic segregation of chromosomes was analysed in three reciprocal translocation carriers, using FISH on interphase spermatozoa. The segregation pattern was first studied in 27,844 spermatozoa from two siblings carrying the reciprocal translocation t(6;11)(q14;p14). Three centromeric probes, specific for chromosomes 6, 11 and 1, were simultaneously hybridized so that all centric fragments as well as the ploidy of each cell could be determined by three colour FISH. For both subjects, the respective frequencies of alternate/adjacent 1, adjacent 2, 3:1 and 4:0 segregation modes were 88%, 9%, 3%and < l%. In another reciprocal translocation t(2; 14)(p23.1;q31), a two colour FISH analysis was performed on 4,610 spermatozoa, using a chromosome 2 centromeric probe and a YAC probe located on the centric fragment of chromosome 14. Frequencies of alternate/adjacent 1, adjacent 2, and 3:1 segregations were 89%, 5.2%, and 5.8% respectively. The segregation of chromosomes X, Y and 1 were also analyzed with three colour FISH on the spermatozoa from all three translocation carriers, in order to detect an interchromosomal effect. Aneuploidy rates for the X and Y chromosomes were found to be in the same range in the three translocation carriers and control donors, but disomy 1 rates were slightly increased in the translocation carriers.
Assignment of the uteroferrin gene (ACP5) to swine chromosome 2q12→q21 by fluorescence in situ hybridization
Authors: Yasue H. , Kusumoto H. , Mikami H.
Published: May 2008

A cosmid clone containing the uteroferrin gene (ACP5) was selected from a cosmid library of swine genomic fragments by colony hybridization using uteroferrin cDNA as a probe. The genomic fragment thus cloned was examined by Southern blot and sequence analysis which demonstrated that it contained at least a part of the uteroferrin gene. The cosmid clone DNA was labeled with biotin, and used as a probe for in situ hybridization to swine chromosomes. Hybridization was visualized by the FITC-labeled streptavidin/biotinylated anti-streptavidin system together with R-banding of chromosomes. The hybridization signals revealed that the uteroferrin gene (ACP5) is located on swine chromosome 2q12→q21.
Mapping of human inter-alpha-trypsin inhibitor family heavy chain-related protein gene (ITIHL1) to human chromosome 3p21→p14
Authors: Tobe T. , Saguchi K. , Hashimoto K. , Miura N.-H. , Tomita M. , Li F. , Wang Y. , Minoshima S. , Shimizu N.
Published: May 2008

Inter-α-trypsin inhibitor family heavy chain-related protein (IHRP) is a novel glycoprotein isolated from human plasma. The cDNA encoding IHRP has recently been cloned from human liver cDNA libraries. We report the mapping of this gene (ITIHL1) by fluorescence in situ hybridization using a 2.5-kb cDNA fragment as a probe. ITIHL1 was localized to chromosome region 3p21→p 14 where the genes of heavy chain 1 and 3 of inter-α-trypsin inhibitor are located. This result, together with significant homology between the nucleotide sequences of ITIHL1 and the heavy chain genes, supports ITIHL1 as being a member of an evolutionary related gene family of ITI heavy chains. Northern blot analysis indicated that IHRP was predominantly synthesized in liver. From Southern blot analysis, it was tentatively concluded that ITIHL1 is a single copy gene.
The EcoRI centromeric satellite DNA of the Sparidae family (Pisces, Perciformes) contains a sequence motive common to other vertebrate centromeric satellite DNAs
Authors: Garrido-Ramos M.A. , Jamilena M. , Lozano R. , Ruiz Rejón C. , Ruiz Rejón M.
Published: May 2008

By means of cloning, sequencing, and fluorescence in situ hybridization, we have determined that the EcoRI satellite DNA family is conserved in the 10 sparid species analyzed here. Its conservation, its chromosomal location at the centromere of each chromosome, and its structural features could make this satellite DNA family an important structural and/or functional element of the centromeres of these species. Monomeric units of this satellite DNA have a consensus length of 187 bp. Its sequence is characterized by a high AT content and the presence of short runs of consecutive AT base pairs. These monomeric EcoRl repeats also contain three to four copies, depending on the species, of a short sequence reflecting the repetitive duplication and subsequent divergence of an ancestral 9-bp sequence in this family. This sequence motive is conserved in some parts of the monomeric units of the different species studied at the same positions, and, precisely, surrounding the area in which the curvature of the monomeric molecule is greatest. The 9-bp sequence motive is similar to other direct-repeat sequences of the centromeric satellite DNAs of other vertebrates, including those of amphibians and mammals.