Assignment of the tenascin gene (HXB) to swine chromosome 1q21.1→q21.3 by fluorescence in situ hybridization

Awata, Takashi; Yamakuchi, Hiroshi; Kumagai, Midori; Yasue, Hiroshi

doi:10.1159/000133931

Cited by 21 publications

(19 citation statements)

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“…The resulting mixture (20 µl) was incubated for 15 min at 37°C to reduce the hybridization signals of repetitive sequences, before applying to chromosome spreads, then with chromosome spreads for 16 hrs at 37°C to hybridize the probe DNA to chromosomal DNA. After hybridization, the spreads were rinsed and processed for detection of the hybridization signals on R-banded chromosomes as described previously [2].…”

Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%

Cloning and Chromosomal Assignment of the Porcine Interleukin-2 Receptor Alpha(IL-2R.ALPHA.) Gene.

Kokuho

Hiraiwa²,

Yasue

et al. 2000

The Journal of Veterinary Medical Science

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ABSTRACT. Porcine genomic DNA encoding a 55 kDa subunit of interleukin-2 receptor (IL-2R), which is termed α chain (IL-2Rα), was cloned by repeated plaque hybridization using IL-2Rα cDNA as a probe. Two different lambda phage clones, one of which encoded exon 1 and the 5'-upstream flanking region of IL-2Rα gene and another encoded the sequence from exon 2 to exon 8, were isolated. By analysis of the 5'-upstream region of the gene, putative binding motifs for transcription factors such as GATA family proteins, Ikaros, NF-κB, NF-IL2Rα and SRF, were found as described in human, murine and bovine genes. Two additional motifs for STAT4 binding were also found in this region. Moreover, using the FISH technique, we assigned the porcine IL-2Rα locus to the distal end of the long arm of chromosome 10 (10q6-qter) where the vimentin gene had been assigned nearby.KEY WORDS: genome mapping, interleukin(IL)-2 receptor, swine.

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Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%

Cloning and Chromosomal Assignment of the Porcine Interleukin-2 Receptor Alpha(IL-2R.ALPHA.) Gene.

Kokuho

Hiraiwa²,

Yasue

et al. 2000

The Journal of Veterinary Medical Science

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“…Fluorescence in situ hybridization: Peripheral blood cells from boars (Large White) were cultured and labeled with 5-bromodeoxyuridine (BrdU), according to the method described previously [3]. The cultured cells were treated with hypotonic solution and fixative, followed by spreading on glass-slides as described previously [22].…”

Section: Mannheimmentioning

confidence: 99%

Section: Mannheimmentioning

confidence: 99%

“…59(6): 431-435, 1997 Mannheim. DNA sequence was determined by dideoxy chain terminator method using PRISM TM ready reaction dyeterminator cycle sequencing kit (Perkin Elmer, U.S.A.) and an ABI 373A DNA sequencer (Perkin Elmer, U.S.A.).Fluorescence in situ hybridization: Peripheral blood cells from boars (Large White) were cultured and labeled with 5-bromodeoxyuridine (BrdU), according to the method described previously [3]. The cultured cells were treated with hypotonic solution and fixative, followed by spreading on glass-slides as described previously [22].…”

mentioning

confidence: 99%

“…For the probe, the DNA of the cosmid clone was labeled with biotin-16-dUTP using a nick translation kit (Boehringer Mannheim, FRG). After hybridization, the spreads were rinsed and processed for detection of the hybridization signals on R-banded chromosomes as described previously [3].For precise localization of the signals on chromosomes, the hybridization signals were collected by cooled CCD camera (Hamamatsu Photonics Co., Japan) installed in a microscope under B2 excitation. The same frame of Rband patterns of the chromosome was then collected by the CCD under G-excitation.…”

mentioning

confidence: 99%

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Structural Organization and Chromosomal Assignment of the Swine Endothelin-1 Gene.

Kojima

Ohata

Yasue

1997

The Journal of Veterinary Medical Science

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ABSTRACT. Cosmid clone containing swine endothelin-1 (EDN1) gene, cosEDN1, was isolated from swine cosmid library using swine EDN1 cDNA as a probe. The sequence analysis of cosEDN1 DNA revealed that the swine EDN1 gene consists of 5 exons, spanning approximately 6.5 kb. In the 5'-upstream region of the EDN1 gene, AP-1 and NF-1 elements were found, suggesting the possibility that the expression of swine EDN1 gene is controlled by protooncogene products Fos and Jun, and TGF-β. Fluorescence in situ hybridization (FISH) using cosEDN1 DNA as a probe demonstrated that EDN1 gene resides on swine chromosome 7p13->pter. -KEY WORDS: EDN1, FISH, gene structure, mapping, swine.J. Vet. Med. Sci. 59(6): 431-435, 1997 Mannheim. DNA sequence was determined by dideoxy chain terminator method using PRISM TM ready reaction dyeterminator cycle sequencing kit (Perkin Elmer, U.S.A.) and an ABI 373A DNA sequencer (Perkin Elmer, U.S.A.).Fluorescence in situ hybridization: Peripheral blood cells from boars (Large White) were cultured and labeled with 5-bromodeoxyuridine (BrdU), according to the method described previously [3]. The cultured cells were treated with hypotonic solution and fixative, followed by spreading on glass-slides as described previously [22].The chromosome spreads were treated with 100 µg/ml of RNase A, then with 70% formamide-2 × SSC to denature the chromosomal DNA according to the method of Péquignot [16]. The resulting spreads were subjected to in situ hybridization under the following conditions; 20 µl of hybridization mixture containing 200 ng of heat denatured probe, 40 µg of denatured Cot5 DNA (repetitive sequenceenriched DNA fraction), 2 × SSC, 50 mM sodium phosphate, 1 × Denhaldt's solution, 10% dextran sulfate, 0.1% SDS and 50% formamide was laid on a spread with a cover slip, followed by incubation for 16 hr at 37°C. For the probe, the DNA of the cosmid clone was labeled with biotin-16-dUTP using a nick translation kit (Boehringer Mannheim, FRG). After hybridization, the spreads were rinsed and processed for detection of the hybridization signals on R-banded chromosomes as described previously [3].For precise localization of the signals on chromosomes, the hybridization signals were collected by cooled CCD camera (Hamamatsu Photonics Co., Japan) installed in a microscope under B2 excitation. The same frame of Rband patterns of the chromosome was then collected by the CCD under G-excitation. The image of hybridization signals was superimposed on that of R-band pattern using image analyzer (IBAS; Carl Zeiss, FRG) to locate signal positions on chromosomes.Endothelin-1 (EDN1) consisting of 21 amino acid residues, which is processed from preproendothelin-1 via intermediate protein called "big endothelin-1", was originally identified as a potent vasoconstrictor peptide from a culture supernatant of porcine aortic endothelial cells [20]. Then the EDN1 has been demonstrated to possess a wide variety of biological activities, including contraction of nonvascular smooth muscles, stimulation of proliferation of va...

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Directed isolation and mapping of microsatellites from swine Chromosome 1q telomeric region through microdissection and RH mapping

et al. 2001

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Several quantitative trait loci (QTLs) (vertebrate number, birth weight, age at puberty, growth rate, gestation length, and backfat depth) have been independently mapped to the distal region of swine Chromosome (SSC) 1q in several resource populations. In order to improve the map resolution and refine these QTLs more precisely on SSC1q, we have isolated and mapped additional microsatellites (ms), using chromosome microdissection and radiation hybrid (RH) mapping. Five copies of the telomeric region of SSC1q were microdissected from metaphase spreads and pooled. The chromosomal fragment DNA was randomly amplified by using degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR), enriched for ms, and subcloned into a PCR vector. Screening of subsequent clones with ms probes identified 23 unique ms sequences. Fifteen of these (65%) were subjected to radiation hybrid (RH) mapping by using the INRA-University of Minnesota porcine RH panel (IMpRH); and the remaining eight were not suited for the RH mapping. Twelve microsatellites were assigned to SSC1q telomeric region of IMpRH map (LOD >6), and three remain unlinked (LOD <6). Out of the 15 microsatellite markers, 9 were polymorphic in NIAI reference population based on the Meishan and Göttingen miniature pig. In summary, we have used microdissection and radiation hybrid mapping to clone and map 12 new microsatellites to the swine gene map to increase the resolution of SSC1q in the region of known QTLs.

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Assignment of the tenascin gene (HXB) to swine chromosome 1q21.1→q21.3 by fluorescence in situ hybridization

Cited by 21 publications

References 2 publications

Cloning and Chromosomal Assignment of the Porcine Interleukin-2 Receptor Alpha(IL-2R.ALPHA.) Gene.

Cloning and Chromosomal Assignment of the Porcine Interleukin-2 Receptor Alpha(IL-2R.ALPHA.) Gene.

Structural Organization and Chromosomal Assignment of the Swine Endothelin-1 Gene.

Directed isolation and mapping of microsatellites from swine Chromosome 1q telomeric region through microdissection and RH mapping

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