• 제목/요약/키워드: Metaphase analysis

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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.

rDNA와 말단소체 반복서열 탐침을 이용한 천마의 FISH 염색체 조성 분석 (Analysis of Chromosome Composition of Gastrodia elata Blume by Fluorescent in situ Hybridization using rDNA and Telomeric Repeat Probes)

  • ;박응준;김현희
    • 한국약용작물학회지
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    • 제26권2호
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    • pp.113-118
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    • 2018
  • Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

인체말초혈액 림프구와 마우스골수세포에서 중기염색체 분석법과 미소핵검사법을 이용한 방사선적응반응 평가 (Cytogenetic Radiation Adaptive Response Assessed by Metaphase Analysis and Micronuclei Test in Human Lymphocytes and Mouse Bone Marrow Cells)

  • 민정준;범희승;이승연;최근희;정환정;송호천;김지열
    • 대한핵의학회지
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    • 제32권6호
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    • pp.525-533
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    • 1998
  • 목적: 인체 말초혈액 림프구와 마우스의 골수세포에서 중기염색체 분석법과 미소핵 검사법을 각각 실시하여 저선량 방사선에 의한 적응반응이 몇 시간째에 가장 유의하게 나타나는지 알아보고, 중기염색체분석법과 미소핵 검사법간에 서로 상관관계를 분석하였다. 대상 및 방법: 건강한 비흡연자의 말초혈액과 ICR계 수컷 마우스 56마리를 대상으로 하였고, Cs-137 조사기를 이용하여 방사선을 조사하였다. 인체 말초혈액 림프구의 중기염색체 분석법은 세포 100개당 불안정 염색체인 반지형과 이 중 중심체형염색체의 숫자를 계수하였으며, 미소핵 검사법은 이핵세포 1,000개에서 나타나는 미소핵의 수를 계수하였다. 마우스 골수세포는 마우스의 대퇴골에서 추출한 후 중기염색체 분석법에 의해 불안정 염색체인 반지형과 이 중 중심체형 염색체의 숫자를 같은 방법으로 계수하였으며, 미소핵 검사법은 마우스 골수세포의 미성숙적혈구 1,000개에서 나타나는 미소핵의 수를 계수하였다. 대조군은 방사선을 조사하지 않은 군이며, 실험군은 0.18 Gy 및 2 Gy 단일조사군, 0.18 Gy 조사 후 4, 7, 12, 24시간 후에 다시 2Gy를 조사한 군으로 나누었고 각각의 군에서 두 검사법의 상관관계를 분석하였다. 결과: 중기영색체 분석법과 미소핵 검사법 모두에서 저선량방사선을 조사한 후 7시간째에 고선량을 조사한 군에 적응반응이 가장 유의하게 나타남을 알 수 있었다. 또한 중기염색체 분석법과 미소핵검사법은 인체 말초혈액림프구를 이용한 실험(r=0.98, p=0.001)과 마우스골수세포를 이용한 실험(r=0.99, p<0.001) 모두에서 매우 높은 상관관계를 보였다. 결론: 방사선에 대한적응반응이 생체내, 외 실험을 통하여 동신에 확증되었고, 저선량 조사 후 4시간 이후에 시작되어 7시간째에 가장 높은 효과를 나타내는 것으로 보인다. 또한 중기염색체 분석법과 미소핵 검사법은 인체 말초혈액 림프구나 마우스 골수세포를 이용한 실험 모두에서 매우 높은 상관관계가 있음을 알 수 있었고, 방사선에 의한 염색체 손상을 분석하고자 할 때 방법이 간편한 미소핵 검사를 사용하여도 중기 염색체분석법과 같은 결과를 얻을 수 있을 것으로 사료되었다.

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Comparison of Sexing Analysis between Karyotyping and Blasomere-PCR in Bovine embryos

  • Chang, Suk-Min;Lee, Jong-Ho;Park, Joong-Hoon;Park, Wha-Sik;Park, Chang-Sik;Jin, Dong-Il
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.92-92
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    • 2003
  • Accurate analysis of nuclear status is needed when biopsied-blastomeres are used for embryo sexing. In this study, the nuclear status of blastomeres derived from 8- to 16-cell stage IVF bovine embryos was analyzed to evaluate the representative of single blastomere for embryo sexing. When 55 embryos were analyzed by PCR following biopsy, the coincident rate of sex determination between biopsied-single blastomere and matched blastocyst by PCR was 80 %. Karyotyping of biastomeres in 8- 16-cell stage bovine embryos was conducted to assess chromosome status of IVF embryos. To establish karyotyping of blastomeres, concentrations of vinblastine sulfate and duration of exposure time for metaphase plate induction with 8- to 16-cell stage bovine embryos were tested. The most effective condition for induction of metaphase plate (>45%) was 1.0 ug/ml vinblastine sulfate treatment for 15 h. In 22 embryos under the condition, only 8 embryos out of ten that had a normal diploid chromosome complement showed a sex-chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four out of the other 11 embryos having a mixoploid chromosomal complement contained haploid blastomere with wrong sex chromosome (18.2%). These results suggested that morphologically normal bovine embryos derived from IVF had considerable proportion of mixoploid and sex-chromosomal mosaicism which could be the cause of discrepancies of the sex between biopsied-single blastomere and matched blastocyst by PCR analysis.

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Frequency of Chromosomal Abnormalities in Pakistani Adults with Acute Lymphoblastic Leukemia

  • Shaikh, Muhammad Shariq;Adil, Salman Naseem;Shaikh, Mohammad Usman;Khurshid, Mohammad
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권21호
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    • pp.9495-9498
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    • 2014
  • Background: The difference in prognosis of adult and childhood acute lymphoblastic leukemia (ALL) can be attributed largely to variation in cytogenetic abnormalities with age groups. Cytogenetic analysis in acute leukemia is now routinely used to assist patient management, particularly in terms of diagnosis, disease monitoring, prognosis and risk stratification. Knowing about cytogenetic profile at the time of diagnosis is important in order to take critical decisions in management of the patients. Aim and Objectives: To determine the frequency of cytogenetic abnormalities in Pakistani adult patients with ALL in order to have insights regarding behavior of the disease. Materials and Methods: A retrospective analysis of all the cases of ALL (${\geq}15$years old) diagnosed at Aga Khan University from January 2006 to June 2014 was performed. Phenotype (B/T lineage) was confirmed in all cases by flow cytometry. Cytogenetic analysis was made for all cases using the trypsin-Giemsa banding technique. Karyotypes were interpreted using the International System for Human Cytogenetic Nomenclature (ISCN) criteria. Results: A total of 166 patients were diagnosed as ALL during the study period, of which 151 samples successfully yielded metaphase chromosomes. The male to female ratio was 3.4:1. The majority (n=120, 72.3%) had a B-cell phenotype. A normal karyotype was present in 51% (n=77) of the cases whereas 49% (n=74) had an abnormal karyotype. Of the abnormal cases, 10% showed Philadelphia chromosome; t(9;22)(q34;q11.2). Other poor prognostic cytogenetic subgroups were t(4;11)(q21;q23), hypodiploidy (35-45 chromosomes) and complex karyotype. Hyperdiploidy (47-57 chromosomes) occurred in 6.6%; all of whom were younger than 30 years. Conclusions: This study showed a relatively low prevalence of Philadelphia chromosome in Pakistani adults with ALL with an increase in frequency with age (p=0.003). The cumulative prevalence of Philadelphianegative poor cytogenetic aberrations in different age groups was not significant (p=0.6).

지식 베이스를 이용한 교육용 염색체 분석 시스템 (Chromosome Analysis System based on Knowledge Base for CAI)

  • 박정선;신용원
    • 한국지능정보시스템학회:학술대회논문집
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    • 한국지능정보시스템학회 2001년도 춘계정기학술대회
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    • pp.215-222
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    • 2001
  • The task for chromosome analysis and diagnosis by experienced cytogenetists are being concerned as repetitive, time consuming job and expensive. FOr that reason, chromosome analysis system based on knowledge base for CAI had been established to be able to analyze chromosomes and obtain necessary advises from the knowledge base instead of human experts. That s to say, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes, and then the inference results by knowledge base could enter the inference data into the database. Experimental data were composed of normal chromosome of 2,736 patients'cases and abnormal chromosomes of 259 patients'cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples. The complete system provides variously morphological information by analysis of normal or abnormal chromosomes and it also has the advantage of being able to consult with user on chromosome analysis and diagnosis.

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Constructing intelligent agent for chromosome knowledge base

  • Shin, Yong-Won
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2003년도 Proceeding
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    • pp.3-9
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    • 2003
  • The task for chromosome analysis and diagnosis by experienced cytogenetists are being concerned as repetitive, time consuming job and expensive. For that reason, intelligent agent based on chromosome knowledge base has been established to be able to analyze chromosomes and obtain necessary advises from the knowledge base instead of human experts. That is to say, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes, and then the inference results by knowledge base could enter the inference data into the database. Experimental data were composed of normal chromosomes of 2,736 patients 'cases and abnormal chromosomes of 259 patients' cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples. The completed intelligent agent for chromosome knowledge base provides variously morphological information by analysis of normal or abnormal chromosomes and it also has the advantage of being able to consult with user on chromosome analysis and diagnosis.

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한우 체외 수정란의 체외 배양 조건에 따른 염색체 분석 (Chromosomal Analysis of Hanwoo Embryos by In Vitro Culture Condition)

  • 최선호;조상래;한만희;김현종;최창용;손동수;정연길;김상근;손시환
    • 한국수정란이식학회지
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    • 제22권2호
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    • pp.137-141
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    • 2007
  • 항산화제는 산소의 저장고로서 무혈청 배양액에서 주요한 작용을 하며, 복합배지에서 유용한 첨가제로 알려져 있다. 따라서 본 연구는 한우 체외 수정란의 배양에 있어서 항산화제인 L-cysteine의 작용과 수정란의 발달 단계별 염색체의 분석을 통하여 체외 수정란의 배양 체계를 수립하고자 실시하였다. 한우 난포란의 체외 성숙은 0.1% PVA, 0.1 mM L-cysteine 첨가 시 체외 성숙율은 73.4%, 94.6%으로 각각 나타냈으며, 처리간에 유의적인 차이를 보였다(p<0.05). 체외 발달율은 20.3%, 10.0%로 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA 첨가구에서 각각 나타났으며, 처리간에 유의적인 차이는 보이지 않았다. 배양액의 종류에 따른 염색체 분석 결과는 중기상의 수정란은 18.3%, 12.0%를 보였으며, 분석 가능 수정란 수는 6.1%, 4.0%로 5% FBS+TCM199, 0.1mM L-cysteine 첨가구에서 각각 나타났고, 60, XX 2개, 60XY 1개가 5% FBS+TCM199 처리구에서, 60, XX 2개가 0.1 mM L-cysteine 처리구에서 확인되었고, 처리간에 유의적인 차이가 없었다. 수정란의 발달 단계별 염색체 분석 결과는 $4{\sim}16$세포기는 5% FBS-TCM199 배양액과 0.1 mM L-cysteine을 첨가한 배양액에서 18.3, 12.0%의 염색체 중기상을 확인할 수 있었고, 상실기에서는 43.1, 13.0%의 염색체 중기상을 보였으며, 배반포기의 경우는 94.8, 100.0%의 염색체 중기상을 보여 발달 단계가 진행될수록 염색체 중기 상이 많이 나타났다. 이상의 결과로 항산화제인 L-cysteine은 한우 난포란의 체외 성숙 및 발달에 중요한 인자임을 확인하였다.

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.

담배나방(Helicoverpa assulta: 인시목 밤나방과)의 정자형성 과정과 염색체수 (Spermatogenesis and Chromosome Number of the Tobacco Budworm Helicoverpa assulta Guenee (Lepidoptera: Noctuidae))

  • 유종명;정성은;박희윤;채순용;김상석
    • 한국연초학회지
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    • 제18권1호
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    • pp.12-20
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    • 1996
  • The spermatogenesis and chromosome number were investigated in the pupal testes of Helicouerpa assulta Guenee by light microscopy. During the spermatogenesis, each bundle of P8(256) sperms developed by 6 mitotic and 2 meiotic spermatogonial divisions. From the early stage of spermatogenesis, it was distinguishable between two kinds of sperm differentiation, eupyrene and apyrene spermatogenesis, which are characteristic in Lepidoptera, by the differences in nuclear shape and cell distribution in immature spermatocyst. Through the followed spermiogenesis, the spermatocysts were developed into two kinds of mature cyst, a streamline-shaped eupyrene cyst with nucleated sperms of thready head or a long spindle-shaped apyrene cyst with anucleated sperms of cylindrical head. As the results off chromosomal analysis at metaphase of the spermatogonial mitosis and spermatocytic meiosis, the chromosome number were 2n=6a/n=31, respectively, and no variation between individuals.

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