• 제목/요약/키워드: Fluorescence in-situ hybridization (FISH)

검색결과 168건 처리시간 0.041초

Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

양적형광접합보인법(Q-FISH)에 의한 돼지 백혈구 세포의 텔로미어 함량 분석 (Amount of Telomeric DNA on Pig Lymphocytes by Quantitative Fluorescence in situ Hybridization)

  • 손시환;정현진;최덕순
    • Journal of Animal Science and Technology
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    • 제50권4호
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    • pp.465-474
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    • 2008
  • 연구에서는 돼지의 세포 분화 및 활성적 특이성과 개체의 성장, 노화의 생물학적 특성을 구명하고자 국내 대표적인 종돈을 대상으로 telomeric DNA probe를 이용한 양적형광접합보인법(Q-FISH)으로서 백혈구 세포의 연령 별, 품종 별, 성 별 텔로미어 함량을 분석 고찰하고, 텔로미어 함유율과 개체 별 각 생산 능력과의 연관성을 검토하고자 하였다. 본 연구에 공시된 품종은 대한양돈협회 제2종돈능력검정소에서 검정 사육된 Yorkshire, Landrace, Duroc 및 Berkshire종 암수 100두 이상으로서 분석 결과, 돼지의 연령이 증가함에 따라 유의한 텔로미어 함유율의 감소를 나타내었고, 품종 간에 있어서도 유색 종이 백색 종에 비해 유의적으로 높은 텔로미어 함유율을 보였으며, 성 간에도 수컷이 암컷에 비해 높은 텔로미어 함유율을 나타내었다. 한편 각 개체 별 능력검정 성적과 텔로미어 함유율 간의 상관관계를 분석한 바 일당 증체량, 정육율 및 선발지수와는 정(+)의 상관을 보이고, 체장, 사료 요구율 및 등지방 두께와는 부(-)의 상관을 보여 선발 지표로서 매우 바람직한 양상을 보이나 이들 간의 상관계수가 낮고 또한 모든 계수가 유의성이 없는 것으로 분석되어 각 경제형질과 개체의 텔로미어 함유율 간에 큰 연관성이 없는 것으로 나타났다. 이상의 결과들을 종합하여 볼 때 텔로미어 함유율이 돼지의 생리 표지로서 활용은 가능하나 생산능력의 표지로서 이용하기에는 다소 어려움이 있는 것으로 사료된다.

체세포분열과 감수분열 및 bicolor FISH를 이용한 섬시호의 세포유전학적 분석 (Cytogenetic Analysis Using Mitosis, Meiosis Chromosomes and bicolor Fluorescence in situ Hybridization of Bupleurum latissimum Nakai)

  • 김수영;방재욱;이중구
    • 한국약용작물학회지
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    • 제14권6호
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    • pp.354-359
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    • 2006
  • Chromosome analysis using mitosis, meiosis and bicolor FISH were carried out in Bupleurum latissimum Nakai, which is one of the endemic plants in Ulleung island of korea. The somatic methaphase chromosomes number of this plant was 2n = 2x = 16 and the chromosome complements consisted of six pairs of metacentrics and two pairs of submetacentrics. The size of chromosomes ranged 2.40${\sim}$4.20 ${\mu}$m and NOR (nucleolus organizer region) chromosome did not observed using conventional staining. In meiosis chromosomes, metaphase-I and anaphase-I were observed. Metaphase-I anaphase-I showed 8 bivalents and chromosomes migration to make two daughter cells. Using bicolor FISH, one pair of 5S and 45S rDNA signals were detected on the centromeric region of chromosome 3 and the end of short of chromosome 2,respectively. We also observed the NOR using 45S rDNA probe.

Circulating Aneuploid Cells Detected in the Blood of Patients with Infectious Lung Diseases

  • Kim, Hongsun;Cho, Jong Ho;Sonn, Chung-Hee;Kim, Jae-Won;Choi, Yul;Lee, Jinseon;Kim, Jhingook
    • Journal of Chest Surgery
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    • 제50권2호
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    • pp.126-129
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    • 2017
  • The identification of circulating tumor cells (CTCs) is clinically important for diagnosing cancer. We have previously developed a size-based filtration platform followed by epithelial cell adhesion molecule immunofluorescence staining for detecting CTCs. To characterize CTCs independently of cell surface protein expression, we incorporated a chromosomal fluorescence in situ hybridization (FISH) assay to detect abnormal copy numbers of chromosomes in cells collected from peripheral blood samples by the size-based filtration platform. Aneuploid cells were detected in the peripheral blood of patients with lung cancer. Unexpectedly, aneuploid cells were also detected in the control group, which consisted of peripheral blood samples from patients with benign lung diseases, such as empyema necessitatis and non-tuberculous mycobacterial lung disease. These findings suggest that chromosomal abnormalities are observed not only in tumor cells, but also in benign infectious diseases. Thus, our findings present new considerations and bring into light the possibility of false positives when using FISH for cancer diagnosis.

Screening for down syndrome using trophoblast retrieval and isolation of the cervix: preliminary study

  • Lee, Min Jin;Kim, Soo Hyun;Park, Hee Jin;Shim, Sung Han;Jang, Hee Yeon;Cha, Dong Hyun
    • Journal of Genetic Medicine
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    • 제17권2호
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    • pp.68-72
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    • 2020
  • Purpose: Trisomy 21, the cause of Down syndrome (DS) with various medical problems, is the most common aneuploidy during the fetal period. For diagnosis, a non-invasive screening test using maternal blood, which cannot be confirmed and invasive confirmation test with a risk of miscarriage, may be performed. The trophoblast retrieval and isolation of the cervix (TRIC) have been proposed by some researchers as an alternative to overcome the limitations of current tests. We experimented using TRIC to identify the possibility of trisomy 21 for the first time in Asia. Materials and Methods: Three cases of DS were analyzed confirmed by invasive tests (chorionic villus sampling, amniocentesis). All samples of trophoblasts immediately were immersed in phosphate-buffered saline and processed with formalin for fixation. The trophoblasts were isolated using an anti-human leukocyte antigen-G antibody coupled to magnetic nanoparticles. β-human chorionic gonadotropin (hCG)-expressing cells were considered as trophoblast cells, and the detection rate calculated. DS was confirmed by fluorescence in situ hybridization (FISH). Results: The mean trophoblast detection rate using β-hCG was 78.1%, and the detection rate using FISH was 22.2%. In all cases, the trisomy of chromosome 21 was identified. Conclusion: Trophoblast can be obtained from the five weeks of gestation and has a high detection rate, so it is noted that it can replace the current prenatal genetic test. To realize the clinical application as a prenatal genetic test, we will need additional efforts to identify trisomy 21 as well as other chromosomal abnormalities in future large-scale studies.

형광직접보합법을 이용한 미배양 양수세포에서 산전 이수배수체 확인 (Prenatal Aneuploidy Detection in Uncultured Amniotic Fluid Interphase Cells by Fluorescence in situ Hybridization (FISH))

  • 설혜원;고희정;송남희;김숙령;이화진;오선경;박중신;전종관;윤보현;신희철;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제30권3호
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    • pp.223-231
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    • 2003
  • Objective: The aim of the present study was to evaluate the clinical efficiency of fluorescent in situ hybridization (FISH) in the prenatal diagnosis of chromosomal aneuploidy. Methods: We reviewed data of 268 cases to identify women undergoing genetic amniocentesis at cytogenetic laboratory, from January 2000 to December 2002. Amniotic fluid was submitted for both rapid FISH on uncultured interphase amniocytes using a commercially available DNA probe for chromosome 13, 18, 21, X, Y and standard karyotyping on cultured metaphase amniocytes. Results from FISH and full karyotype were compared. Results: There were 251 cases (84%) normal and 17 cases (16%) abnormal in FISH results. All 17 cases of trisomy 13, 18, 21 including two cases of mosaicism and sex chromosome aneuploidies which are detected by FISH were confirmed with conventional cytogenetics and there was no false positive result. Twenty two cases had karyotypically proven abnormalities that could not have been detected by the targeted FISH. Conclusion: Interphase FISH analysis of uncultured amniotic fluid cells has been shown to be an effective and reliable technique for rapid fetal aneuploidy screening during pregnancy as an adjunctive test to conventional cytogenetics.

Streptococcus mutans의 gtfD 유전자 발현에 대한 이온 및 완충액의 영향 (THE EFFECTS OF IONS AND BUFFER SOLUTIONS ON THE MRNA EXPRESSION OF gtfD GENE OF Streptococcus mutans)

  • 김보영;김신;정진
    • 대한소아치과학회지
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    • 제31권2호
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    • pp.314-322
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    • 2004
  • 글루캔 형성은 이온, 완충액 및 영양물질 등과 같은 구강 내 존재하는 물질과 이들의 변화에 영향을 받는다. 본 연구에서는 치아 우식증의 중요한 원인균인 Streptococcus mutans를 실험균으로 하여 수용성 글루캔 합성 효소인 GTFD 유전자의 발현에 대한 이온 및 완충액의 영향을 Fluorescent in situ hybridization 방법으로 관찰하여 다음의 결과를 얻었다. 1. $CaCl_2$ 농도가 1.0mM, KCl 농도가 2.5mM, $MgCl_2$의 농도가 0.4mM일 때 생균수가 가장 많았다. 2. 완충액의 경우 sodium bicarbonate 10mM, sodium phosphate 1mM, potassium phosphate 0.1mM에서 생균수가 가장 많았고, 3가지 완충액 모두 100mM 농도에서 생균수가 가장 적었다. 3. 자당이 함유되지 않은 BHI 배지에 비해 1%자당을 첨가한 경우 gtfD 유전자의 mRNA 발현에 따른 녹색형광이 관찰되었다. 4. $CaCl_2$ 0.25mM일 때 gtfD 유전자의 mRNA 발현에 따른 녹색형광이 대조군보다 강하였고, KCl의 경우 10mM일 때 대조군과 유사한 녹색형광을 나타냈으나, 다른 농도에서는 거의 관찰되지 않았다. $MgCl_2$의 경우 각 농도에서 대조군보다 녹색형광이 약하게 나타났다. 5. Sodium bicarbonate, sodium phosphate 완충액의 경우 mRNA 발현에 따른 녹색형광이 각 농도에서 대조군과 유사하게 나타났으며, potassium phosphate 완충액의 경우 100mM에서 녹색형광을 거의 관찰할 수 없었다.

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성장부진과 발달지연을 보인 환아에서 확인된 환상 9번 염색체 1례의 세포유전학적인 연구 (Cytogenetic evaluation of a patient with ring chromosome 9 presenting failure to thrive and developmental delay)

  • 박윤미;노한내;김숙자;안영민
    • Clinical and Experimental Pediatrics
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    • 제51권4호
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    • pp.426-430
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    • 2008
  • 환상 염색체의 발생 기전은 염색체 말단 부분이 결손된 후 양 끝이 융합되거나 끝분절 염기서열이 앞뒤역순상동서열로 융합될 경우로 생각되고 있다. 환상 염색체는 세포 분열을 하는 동안 불안정하기 때문에 세포핵이 없는 딸세포의 사망률이 증가하게 되어 생존하는 세포수가 감소하고 성장 장애가 발생하게 된다. 표현형은 염색체 손실의 정도에 따라 다양하다. 저자들은 최초로 심각한 저신장을 주소로 내원한 환아의 염색체 검사상 환상 9번 염색체를 확인하였기에 이를 보고하는 바이다.

SHARON/ANAMMOX 결합공정에서 슬러지의 입상화와 특성 (Granulation and Characteristics of Sludges in the Combined SHARON/ANAMMOX Processes)

  • 황인수;민경석
    • 한국물환경학회지
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    • 제22권2호
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    • pp.300-307
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    • 2006
  • The combined SHARON (Single reactor system for High ammonium Removal Over Nitrite)-ANAMMOX (Anaerobic ammonium oxidation) reactor were operated in mesophilic condition ($35^{\circ}C$). In this study, microbial granulation and characteristics of SHARON and ANAMMOX sludges were investigated using settling test, Scanning Electron Microscopy (SEM) and Fluorescence In Situ Hybridization (FISH). In SHARON reactor, Aerobic granulation with diameter of 1.5~2.5 mm was accomplished but aerobic granulation was weaker than anaerobic granular sludge. Initial seed sludge of ANAMMOX reactor was used as attached media for biofilm growth. ANAMMOX sludge was more compact and rounder rather than seed sludge. Though ANAMMOX sludge has high activity, it has lower settling ability than the seed granule. The color of ANAMMOX sludge was changed from dark to redish brown granular with diameter of 1~2 mm. In FISH of ANAMMOX sludge, high fraction of Candidatus B. stuttgartiensis which paid great role of nitrogen conversion was detected. Also, FISH results reveals that ANAMMOX bacteria inhabit at inner parts near surface, having advantages in utilization of substrates and protection from oxygen inhibition.

Upregulation by KCI Treatment of Eukaryotic Translation Elongation Factor 1A (eEF1A) mRNA in the Dendrites of Cultured Rat Hippocampal Neurons

  • Moon, Il Soo;Cho, Sun-Jung;Lee, HyunSook;Seog, Dae-Hyun;Jung, Yong Wook;Jin, IngNyol;Walikonis, Randall
    • Molecules and Cells
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    • 제25권4호
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    • pp.538-544
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    • 2008
  • Activity-dependent local translation in the dendrites of brain neurons plays an important role in the synapse-specific provision of proteins necessary for strengthening synaptic connections. In this study we carried out combined fluorescence in situ hybridization (FISH) and immunocytochemistry (IC) and showed that more than half of the eukaryotic elongation factor 1A (eEF1A) mRNA clusters overlapped with or were immediately adjacent to clusters of PSD-95, a postsynaptic marker, in the dendrites of cultured rat hippocampal neurons. Treatment of the neurons with KCl increased the density of the dendritic eEF1A mRNA clusters more than two-fold. FISH combined with IC revealed that the KCl treatment increased the density of eEF1A mRNA clusters that overlapped with or were immediately adjacent to PSD-95 clusters. These results indicate that KCl treatment increases both the density of eEF1A mRNA clusters and their synaptic association in dendrites of cultured neurons.