• 제목/요약/키워드: genomic in situ hybridization

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

  • 김철우;이을태;김화영;최인후;방진기;구달회;방재욱
    • 한국육종학회지
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    • 제41권4호
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    • pp.468-473
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    • 2009
  • 본 연구에서는 양파와 파간 종간교잡 F1의 불임성의 한 요인으로 여겨지는 염색체 이상과 교잡식물체의 염색체의 조성을 밝혀 향후 종간교잡을 이용한 양파의 형질개량과정에 종간교잡 육종의 효율성을 높이고자 하였다. 종간교잡식물체의 감수분열 염색체를 관찰한 결과 중기에 7쌍의 2가염색체와 2개의 1가염색체가 관찰되었다. 체세포 염색체수는 16개로 양파, 파의 염색체 수와 같았으며 GISH한 결과 파와 양파의 염색체가 각각 8개씩 확인되었다. 5S, 45S, TSD DNA를 probe로 하여 양파, 파, 종간잡종의 염색체의 signal을 관찰한 결과 5S와 45S rDNA는 는 3종 모두에서 1쌍의 염색체에서 나타났는데 5S의 경우 염색체의 subcentromeric 에서 45S는 부수체 염색체에서 관찰되었다. TSD DNA는 염색체의 telomeric 부위에서 관찰되었으나 양파의 경우 2쌍은 염색체의 말단부와 중심체 부분에서 signal이 관찰되었고 종간교잡 세포에서도 2쌍이 관찰되어 양파에서 유래된 염색체임이 확인되었다.

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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    • 제3권1호
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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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밀 유전 배경에서 호밀 B 염색체가 감수분열 염색체 접합에 미치는 영향 (Effect of Rye B chromosome on Meiotic Chromosome Association in Wheat (Triticum aestivum L.) Genetic Background)

  • 조성우
    • 한국자원식물학회지
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    • 제35권5호
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    • pp.659-666
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    • 2022
  • 이 연구에서는 밀 첨가 계통의 생식세포분열시, 호밀 B 염색체가 염색체 접합에 미치는 영향을 관찰하였다. 밀 첨가 계통은 동조관계에 있는 Leymus mollis와 L. racemosus 염색체를 각각 하나씩 가지고 있다. 밀의 유전적 배경에서 두 Leymus염색체의 이동은 genomic in situ hybridization에 의하여 확인되었다. 호밀 B 염색체를 가지고 있지 않은 밀 첨가 계통의 생식세포 제1중기 분열기에 두 Leymus 염색체의 대부분은 일가염색체를 보였다(98.1 ± 0.5%). 반면에 호밀 B 염색체를 가지고 있는 밀 첨가 계통에서 Leymus 이가염색체의 빈도(10.3 ± 0.2%)는 호밀 B 염색체를 가지고 있지 않은 밀 첨가 계통의 Leymus이가염색체의 빈도(1.9 ± 0.5%)보다 높았다. 호밀 B 염색체를 가지고 있지 않은 밀 첨가 계통에서는 비정상적인 구조를 가지고 있는 이가염색체가 관찰되었다. 반면, 비록 매우 낮은 빈도이지만 호밀 B 염색체를 가지고 있는 밀 첨가 계통에서는 정상적인 형태를 보이는 이가염색체가 확인되었다. 호밀 B 염색체의 영향은 또한 보통밀과 L. racemosus염색체 사이의 이가염색체의 형성을 유도하였으며, 보통밀과 L. mollis 염색체 사이의 삼가염색체의 형성도 유도하였다. 뿐만 아니라 보통밀 염색체 사이에서 초과 교차가 일어나는 것을 확인하였으며, 이러한 현상으로 원, 막대, 그리고 후라이팬 모양의 일반적인 이가염색체의 형태가 아닌 막대모양이나 응축된 형태의 보통밀 이가염색체가 확인되었다.

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

  • 강지언
    • 대한임상검사과학회지
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    • 제52권4호
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    • pp.310-316
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    • 2020
  • 마이크로어레이 진단 기법의 발달은 세포유전학적 관점에서, 다양한 종류의 유전학적 질병과 관련하여 새로운 정보를 제공하고, 질병에 대한 기본적인 통찰력을 제공하는데 매우 중요한 역할을 제공하고 있다. 그동안 많은 연구들에서, 마이크로어레이 기술을 활용한 인간 게놈의 유동성과 다양성을 입증해 주었으며, 게놈의 취약성을 식별하기 위한 보다 정확한 진단기법과 적절한 임상 관리 방법을 효율적으로 제공해 왔다. 앞으로 다양한 유전과 관련된 질병에 기존 세포유전학적 방법을 자동화된 마이크로어레이 방법으로 전환한다면, 보다 효율적인 방법으로 질병을 진단하고, 정확성을 향상시키며, 유전자 배열의 암호화 및 복잡한 특성을 밝히는데 매우 중요한 역할을 할 것으로 생각된다. 또한 이 분석 기법을 활용하여 게놈과 인간의 건강, 질병과의 관계를 분석하여 다양한 정보를 미리 제공하여 질병을 예방하고, 질병의 진단 및 치료에도 도움이 될 수 있는 새로운 혁명을 일으킬 수 있을 것으로 기대된다.

Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • 한국생명과학회:학술대회논문집
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    • 한국생명과학회 2003년도 제40회 국제학술심포지움
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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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    • 제4권5호
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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
    • 원예과학기술지
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    • 제33권1호
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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.

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

  • 전소영;배옥석;김종봉
    • 대한의생명과학회지
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    • 제4권2호
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    • pp.121-128
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    • 1998
  • 본 연구에서는 genomic DNA 전기영동과 TUNEL labeling법을 이용하여 정상대장조직, 대장암조직, 대장암 인접림프절조직과 대장암환자 혈액에서 apoptosis 발현을 분석하였다. 정상대장조직 37례중 4례에서, DNA ladder가 확인되었고, 암조직은 47례중 20례에서, 림프절조직은 15례중 5례에서 나타났으며 대장암환자혈액은 7례 모두에서 발현되지 않았다. TUNEL labeling법을 이용한 조직상의 in situ apoptosis발현은 암조직과 림프절조직에서 확인되었다. 따라서 본 연구에서는 대장암이 진행됨에 따라 apoptosis 발현비가 증가되었으므로 apoptotic index가 대장암 발현과 관련되어 있는 듯 하며 예후예측지표자로서 이용될 가능성이 있다고 사료된다.

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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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    • 제59권2호
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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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    • 제38권3호
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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.