• Title/Summary/Keyword: Genomic in situ hybridization

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Chromosome Analysis Using GISH and FISH of Interspecific Hybrids between Allium cepa L. and A. fistulosum L. (GISH와 FISH를 이용한 양파와 파간 종간교잡계통의 염색체 분석)

  • Kim, Cheol-Woo;Lee, Eul-Tai;Kim, Hwa-young;Choi, In-Hu;Bang, Jin-Ki;Koo, Dal-Hoe;Bang, Jae-Wook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.468-473
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    • 2009
  • Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used for chromosome analysis of hybrids (2n=16) between onion (Allium cepa L., 2n=2X=16) and welsh onion (A. fistulosum L., 2n=2X=16). 5S rDNA, 45S rDNA, and tandemly repeated DNA (TSD) sequence were used as probes for FISH analysis. A. fistulosum specific DNA probe of telomeric repeats and A. fistulosum DNA were used for GISH analysis. In the analysis of meiotic chromosome GISH revealed that hybrids have 7 bivalants and 2 univalents chromosome and 2 univalents were derived from A. fistulosum chromosomes. In somatic chromosomes of hybrid each 8 chromosomes were derived from A. cepa and A. fistulosum, respectively. FISH signal of 45S rDNA probe in A. fistulosum was detected at secondary constriction of chromosomes, while FISH signal in A. cepa was observed in both secondary constriction and telomere of chromosomes. TDS signals in A. fistulosum chromosomes were detected at all subtelomeric of 8 chromosomes and also in 2 pericentromeric of the chromosomes, whereas TDS signals in A. cepa were observed only in subtelomeric in all chromosomes. The pattern of TDS signal in hybrid chromosomes was similar to those of A. fistulosum chromosomes.

Production and Characterizations of Somatic Hybrids between Brassica campestris L. ssp pekinensis and Brassica of oleracea L. var capitata

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.33-38
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    • 2001
  • Protoplasts isolated from inbred lines of Brassica oleracea L. var capitata (cabbage) and Brassica campestris L. ssp. pekinensis (Chinese cabbage) were fused by PEG-mediated method, and somatic hybrid cells were differentiated into plants. for the identification of somatic hybrid plants, ploidy level, plant morphology, and cytological analysis were performed. All of the regenerated plants derived from fused protoplasts were shown to be 2X-4X, or higher ploidy level, presumably due to somatic hybridization or chromosome doubling. The morphology of leaves, petioles, and flowers showed an intermediate phenotype between Chinese cabbage and cabbage. Chromosome numbers in these somatic hybrids ranged mostly from 33 to 38. According to Genomic in situ hybridization (GISH) pattern, signals from both fusion parents of B.campestris or B.oleracea were detected in different colors when chromosomes of putative somatic hybrids were observed.

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Effect of Rye B chromosome on Meiotic Chromosome Association in Wheat (Triticum aestivum L.) Genetic Background (밀 유전 배경에서 호밀 B 염색체가 감수분열 염색체 접합에 미치는 영향)

  • Cho, Seong-Woo
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.659-666
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    • 2022
  • The effect of rye B chromosome (rye B) on chromosome association was investigated in meiosis of wheat addition line. The wheat addition line was with one Leymus mollis chromosome and one L. racemosus chromosome which are under homoeologous relationship. Chromosome behavior of the two Leymus chromosomes in wheat genetic background was revealed by genomic in situ hybridization. In the first metaphase, most of the two Leymus chromosomes showed univalent in the wheat addition line without rye B (98.1 ± 0.5%). On the other hand, the wheat addition line with rye B showed higher frequency of bivalent (10.3 ± 0.2%) than wheat addition line without rye B (1.9 ± 0.5%). The wheat addition line without rye B showed abnormal bivalents with abnormal structure while the wheat addition line with rye B showed normal bivalent in low frequency. By rye B, some bivalent was composed of wheat and L. racemosus, and some trivalent was composed of wheat bivalents with L. mollis chromosome. Also, some wheat bivalents showed hyper-crossover, so those wheat bivalents showed abnormal structure compared to other wheat bivalents with normal structure such as ring, rod, and pan.

Medical Implementation of Microarray Technology (마이크로어레이 분석기법의 임상적용에 관한 연구)

  • Kang, Ji Un
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.310-316
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    • 2020
  • Microarray technology represents a critical new advance in molecular cytogenetics. The development of this approach has provided fundamental insights into the molecular pathogenesis in clinical cytogenetics and has provided a clue to many unidentified or unexplained diseases. The approach allows a comprehensive investigation of thousands and millions of genomic loci simultaneously and enables the efficient detection of copy number alterations. The application of this technology has shown tremendous fluidity and complexity of the human genome, and has provided accurate diagnosis and appropriate clinical management in a timely and efficient manner for identifying genomic alterations. The clinical impact of the genomic alterations identified by microarrays is evolving into a diagnostic tool to identify high-risk patients better and predict patient outcomes from their genomic profiles. The transformation of conventional cytogenetics into an automated discipline will improve diagnostic yield significantly, leading to accurate diagnosis and genetic counseling. This article reviews cytogenetic technologies used to identify human chromosome alterations and highlights the potential utility of present and future genome microarray technology in the diagnosis.

Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • Proceedings of the Korean Society of Life Science Conference
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    • 2003.10a
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    • pp.14-27
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    • 2003
  • The genomes of Arabidopsis and rice have been fully sequenced. Genomic sequencing provides global information about genome structure and organization. A comprehensive research account of our recent studies conducted on genome painting, comparative genomics and genome fusion is provided in order to project the prospects of plant cytogenetic research in post-genomics era. Genome analysis by GISH using genome painting is demonstrated as an excellent means suitable for visualization of a whole genome, since total genomic DNA representing the overall molecular composition of the genome is used as a probe. FISH on extended DNA fibers has been developed for high-resolution FISH and has contributed to determining the copy number and order of genes. We have also mapped a number of genes involving starch synthesis on wheat chromosomes by FISH and compared the position of these genes on linkage map of rice. Macro synteny between wheat and rice can be observed by comparing the location of these genes in spite of the fact that the size of DNA per chromosome differs by 20 fold in two. Moreover, to approach our goal towards making bread and udon noodles from rice flour in future by incorporating bread making and the noodle qualifies in rice, we have been successful in introducing large genomic DNA fragments containing agronomically important genes of wheat into a rice by successive introduction of large insert BAC clones, there by expanding genetic variability in rice. We call this method genome fusion.

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Amperozide Decreases Cocaine-Induced Increase in Behavior and Immediate Early Gene Expression in the Dorsal Striatum

  • Choe, Eun-Sang;Kim, Jong-Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.5
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    • pp.361-367
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    • 2000
  • Cocaine functions as indirect dopamine and serotonin (5-hydroxytryptamine, 5HT) agonists and induces genomic and behavioral alterations in the striatum. Previously we demonstrated that ritanserin, a 5HT2/1C receptor antagonist, is not responsible for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum (see the previous paper in this issue). In this study, it was hypothesized that dopamine and 5HT2/1C receptors are required for cocaine-induced behavioral alterations and c-fos and zif268 mRNA expression. This hypothesis was addressed by infusing amperozide which antagonizes both 5HT2/1C and dopamine receptors and was analyzed using the quantitative in situ hybridization histochemistry in vivo. Systemic injection of amperozide (5 mg/kg, s.c.) significantly blocked increase in behavior, c-fos and zif268 mRNA expression induced by 15 mg/kg cocaine, i.p., in the dorsal striatum. These data suggest that dopamine and 5HT2/1C receptors are necessary for cocaine-induced behavioral alterations and immediate early gene expression in the dorsal striatum.

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FISH Karyotype and GISH Meiotic Pairing Analyses of a Stable Intergeneric Hybrid xBrassicoraphanus Line BB#5

  • Belandres, Hadassah Roa;Waminal, Nomar Espinosa;Hwang, Yoon-Jung;Park, Beom-Seok;Lee, Soo-Seong;Huh, Jin Hoe;Kim, Hyun Hee
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.83-92
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    • 2015
  • xBrassicoraphanus line BB#5, a new synthetic intergeneric hybrid between Brassica rapa L. ssp. pekinensis and Raphanus sativus L. var. rafiphera induced by N-methyl-N-nitroso-urethane mutagenesis in microspore culture, shows high seed fertility and morphological uniformity. Dual-color fluorescence in situ hybridization (FISH) using 5S and 45S rDNA probes and genomic in situ hybridization (GISH) using B. rapa genomic DNA probe were carried out to analyze the chromosome composition and the meiosis pairing pattern compared to its parental lines. The somatic chromosome complement is 2n = 38, which consists of 17 metacentric and two submetacentric chromosomes with lengths of 2.18 to $5.01{\mu}m$. FISH karyotype analysis showed five and eight pairs of 5S and 45S rDNA loci. GISH meiosis pairing analysis showed that 19 complete bivalents were most frequent and accounted for 42% of the 100 pollen mother cells examined. Based on chromosome number, size, morphology, rDNA distribution, and meiosis pairing pattern, both parental genomes of B. rapa and R. sativus appear to exist in xBrassicoraphanus line BB#5, demonstrating its genome integrity. Such stable chromosome constitutions and meiotic pairing patterns in somatic and meiotic cells are very rare in natural and synthetic intergeneric hybrids. Chromosomal studies and genetic and phenotypic changes in allopolyploids a re discussed. The results p resented h erein will b e usef ul f or f urther g enomic s tudy o f xBrassicoraphanus lines and their improvement as promising new breeding varieties.

Prognosis of Colorectal Cancer and Apoptosis Activation (대장암의 예후와 Apoptosis 활성)

  • 전소영;배옥석;김종봉
    • Biomedical Science Letters
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    • v.4 no.2
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    • pp.121-128
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    • 1998
  • We studied on the expression of apoptosis in colorectal cancer, lymph node, their corresponding normal mucosa and colorectal cancer patient's blood by genomic DNA electrophoresis and TUNEL labeling method. From 7 cases among 37, 20 cases among 47 and 5 cases among 15, DNA ladders were expressed in normal tissues, colorectal tissues and Iymph node tissues, respectively. A DNA ladder was not observed in 7 cases of colorectal cancer patients blood. In case of TUNEL labeling, we could observe TUNEL color espression in colorectal cancer and lymph node tissues. As these result suggest that apoptotic index may be associated with the colorectal cancer development, and mat be used as a prognostic indicator but further evaluations will be needed.

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A new mosaic der(18)t(1;18)(q32.1;q21.3) with developmental delay and facial dysmorphism

  • Choi, Young-Jin;Shin, Eunsim;Jo, Tae Sik;Moon, Jin-Hwa;Lee, Se-Min;Kim, Joo-Hwa;Oh, Jae-Won;Kim, Chang-Ryul;Seol, In Joon
    • Clinical and Experimental Pediatrics
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    • v.59 no.2
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    • pp.91-95
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    • 2016
  • We report the case of a 22-month-old boy with a new mosaic partial unbalanced translocation of 1q and 18q. The patient was referred to our Pediatric Department for developmental delay. He showed mild facial dysmorphism, physical growth retardation, a hearing disability, and had a history of patent ductus arteriosus. White matter abnormality on brain magnetic resonance images was also noted. His initial routine chromosomal analysis revealed a normal 46,XY karyotype. In a microarray-based comparative genomic hybridization (aCGH) analysis, subtle copy number changes in 1q32.1-q44 (copy gain) and 18q21.33-18q23 (copy loss) suggested an unbalanced translocation of t(1;18). Repeated chromosomal analysis revealed a low-level mosaic translocation karyotype of 46,XY,der(18)t(1;18) (q32.1;q21.3)[12]/46,XY[152]. Because his parents had normal karyotypes, his translocation was considered to be de novo. The abnormalities observed in aCGH were confirmed by metaphase fluorescent in situ hybridization. We report this patient as a new karyotype presenting developmental delay, facial dysmorphism, cerebral dysmyelination, and other abnormalities.

An update of preimplantation genetic diagnosis in gene diseases, chromosomal translocation, and aneuploidy screening

  • Chang, Li-Jung;Chen, Shee-Uan;Tsai, Yi-Yi;Hung, Chia-Cheng;Fang, Mei-Ya;Su, Yi-Ning;Yang, Yu-Shih
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.3
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    • pp.126-134
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    • 2011
  • Preimplantation genetic diagnosis (PGD) is gradually widely used in prevention of gene diseases and chromosomal abnormalities. Much improvement has been achieved in biopsy technique and molecular diagnosis. Blastocyst biopsy can increase diagnostic accuracy and reduce allele dropout. It is cost-effective and currently plays an important role. Whole genome amplification permits subsequent individual detection of multiple gene loci and screening all 23 pairs of chromosomes. For PGD of chromosomal translocation, fluorescence $in-situ$ hybridization (FISH) is traditionally used, but with technical difficulty. Array comparative genomic hybridization (CGH) can detect translocation and 23 pairs of chromosomes that may replace FISH. Single nucleotide polymorphisms array with haplotyping can further distinguish between normal chromosomes and balanced translocation. PGD may shorten time to conceive and reduce miscarriage for patients with chromosomal translocation. PGD has a potential value for mitochondrial diseases. Preimplantation genetic haplotyping has been applied for unknown mutation sites of single gene disease. Preimplantation genetic screening (PGS) using limited FISH probes in the cleavage-stage embryo did not increase live birth rates for patients with advanced maternal age, unexplained recurrent abortions, and repeated implantation failure. Polar body and blastocyst biopsy may circumvent the problem of mosaicism. PGS using blastocyst biopsy and array CGH is encouraging and merit further studies. Cryopreservation of biopsied blastocysts instead of fresh transfer permits sufficient time for transportation and genetic analysis. Cryopreservation of embryos may avoid ovarian hyperstimulation syndrome and possible suboptimal endometrium.