• 제목/요약/키워드: Fluorescence in situ hybridization analysis

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Comparative Cytogenetic Characteristics and Physical Mapping of the 17S and 5S Ribosomal DNAs between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

  • Bang, Kyong-Hwan;Koo, Dal-Hoe;Kim, Hong-Sig;Song, Beom-Heon;Cho, Yong-Gu;Cho, Joon-Hyeong;Bang, Jae-Wook
    • 한국약용작물학회지
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    • 제11권4호
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    • pp.311-315
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    • 2003
  • This study was carried out to compare chromosomal characteristics between Atractylodes japonica and A macrocephala. Cytogenetic analysis was conducted based on karyotype analysis and physical mapping using fluorescence in situ hybridization. As a result of karyotype analysis by feulgen staining, somatic chromosome numbers of A. japonica and A. macrocephala were 2n=24. The length. of the mitotic metaphase chromosomes of A. japonica ranged from $0.70\;to\;1.60{\mu}m$ with a total length. of $12.11{\mu}m$ and the homologous chromosome complement comprised six metacentrics, five submetacentrics and one subtelocentrics. On the other hand, the length of the mitotic metaphase chromosomes of A. macrocephala ranged from $0.90\;to\;2.35{\mu}m$ with a total length of $16.58{\mu}m$ and the homologous chromosome complement comprised seven metacentrics and five submetacentrics. The total length of A. japonica chromosomes was shorter than that of A. macrocephala, but A. japonica had one subtelocentrics (chromosomes 4) different from A. macrocepha1a. chromosomes. The F1SH technique using 17S and 5S rDNA was applied to metaphase chromosomes. The signals for 17S rDNA were detected on the telomeric regions of chromosomes 4 and 5 in both A japonica and A. macrocephala. The 5S rDNA signal was found in the short arm of chromosome 1.

Identification of a Novel PGE2 Regulated Gene in SNU1 Gastric Cancer Cell

  • Park, Min-Seon;Kim, Hong-Tae;Min, Byung-Re;Kimm, Ku-Chan;Nam, Myeong-Jin
    • BMB Reports
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    • 제33권2호
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    • pp.184-187
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    • 2000
  • Prostaglandin $E_2$ ($PGE_2$) plays an important role in the regulation of various gastric functions, and the growth-inhibitory activities on tumor cells are studied in vitro and in vivo. Although the mechanisms have attracted many researchers in the past decade, the molecular mechanisms of cell cycle arrest, or induction of apoptosis by $PGE_2$, is unclear. We investigated the effects of $PGE_2$ on the growth of the human gastric carcinoma cell line SNU1 and genes that are regulated by $PGE_2$ and isolated them using differential display RT-PCR (DD RT-PCR). FACS analysis suggested that SNU1 cells were arrested at the G1 phase by $PGE_2$ treatment. This growth inhibitory effect was in a time- and dose-dependent manner. Treatment of SNU1 cells with $10\;{\mu}g/ml$ $PGE_2$, followed by DD RT-PCR analysis, revealed differently expressed bands patterns from the control. Among the differently expressed clones, we found an unidentified cDNA clone (HGP-27) overexpressed in $PGE_2$-treated cells. The full-length cDNA of HGP-27 was isolated using RACE, which consisted of a 30-nt 5'-noncoding region, a 891-nt ORF encoding the 296 amino acid protein, and a 738-nt 3'-noncoding region including a poly(a) signal. This gene was localized on the short arm of chromosome number 11. Using the Motif Finder program, a myb-DNA binding repeat signature was detected on the ORF region. The COOH-terminal half was shown to have similarity with the $NH_3$-terminal domain of thioredoxin (Trx). This relation between HGP-27 and Trx implied a potential role for HGP-27 in modulating the DNA binding function of a transcription factor, myb.

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살수여상에서의 질소, 인 제거 미생물 분포 및 질산화 활성 조사 (Analysis on the distribution of nitrogen and phosphorus removing microorganisms and nitrifying activity in a trickling filter)

  • 김동진;유익근;안대희
    • 한국환경과학회지
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    • 제18권6호
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    • pp.691-698
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    • 2009
  • Trickling filter has been extensively studied for the domestic wastewater treatment especially for the small scale plants in rural area. The performance of the trickling filter depends on the microbial community and their activity in the biofilms on the media. Nitrification. denitrification, and phosphorus removal of the trickling filter from the wastewater depend on the activity and the amount of the specific microorganisms responsible for the metabolism. For the estimation of the performance of a trickling filter, batch nitrification experiment and fluorescence in situ hybridization (FISH) were carried out to measure the microbial activity and its distribution on the media of the trickling filter. Batch nitrification activity measurement showed that the top part of the 1st stage trickling filter had the highest nitrification activity and the maximum activity was 0.002 g $NH_4$-N/g MLVSS${\cdot}$h. It is thought that higher substrate (ammonia) concentration yields more nitrifying bacteria in the biofilms. The dominant ammonia oxidizer and nitrite oxidizer in the biofilm were Nitrosomonas species and genus Nitrospira, respectively, by FISH analysis. Less denitrifiers were found than nitrifiers in the biofilm by the probe Rrp1088 which specifically binds to Rhodobacter, Rhodovulum, Roseobacter, and Paracoccus. Phosphorus accumulating bacteria were mostly found at the surface of the biofilm by probe Rc988 and PAO651 which specifically binds to Rhodocyclus group and their biomass was less than that of nitrifiers.

Centromere Repeat DNA Originated from Brassica rapa is Detected in the Centromere Region of Raphanus sativus Chromosomes

  • Hwang, Yoon-Jung;Yu, Hee-Ju;Mun, Jeong-Hwan;Bok, Kwang;Park, Beom-Seok;Lim, Ki-Byung
    • 원예과학기술지
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    • 제30권6호
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    • pp.751-756
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    • 2012
  • Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA sequences in the specific region of the chromosomes. As well as for the integrated physical mapping, FISH karyotype analysis has to be preceded. Karyotype of Raphanus sativus 'Wonkyo 10039' was analyzed by a dual-color FISH technique; using various repetitive DNA probes, including 5S rDNA, 45S rDNA, and centromere retrotransposon. The length of the somatic metaphase chromosome ranged from 1.35 to $2.06{\mu}m$ with a total length of $15.29{\mu}m$. The chromosome complements comprised of eight pairs of metacentrics and one pair of submetacentric. Bleached DAPI Band analysis revealed a heterochromatin region, covering 28.6% to 50.4% each chromosomes. 5S and 45S rDNA sequences were located on two and three pairs of chromosomes, respectively. The centromere retrotransposon of Brassica (CRB) is a major component in Brassica related species that has been maintained as a common centromere component. CRB signals were detected on the centromere and pericentromeric region of R. sativus 'Wonkyo 10039' and three basic Brassica species (B. rapa, B. nigra, and B. oleracea). These results will provide a valuable background for physical mapping and elucidation of the evolutionary relationship among the Brassica related species.

깽깽이풀의 핵형분석과 McFISH를 이용한 rDNA의 물리지도 작성 (Karyotype Analysis and Physical Mapping of rDNAs Using McFISH in Jeffersonia dubia Benth)

  • 김수영;최혜운;구달회;김찬수;방재욱
    • 한국약용작물학회지
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    • 제13권1호
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    • pp.48-51
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    • 2005
  • 보호식물이며, 약용식물인 깽깽이풀 (Jeffersonia dubia)을 대상으로 핵형 분석과 McFISH 기법을 이용한 염색체 분석을 수행하여 다음과 같은 결과를 얻었다. 체세포 염색체 수는 2n=2x=12였으며, 2쌍의 중부 염색체 (염색체 1, 3), 2쌍의 차중부 염색체 (염색체 2, 4) 그리고 2쌍의 차단부 염색체 (염색체 5, 6)로 구분되었고, 염색체의 평균 길이는 $1.95{\sim}3.50{\mu}M$이었다. McFISH기법을 이용하여 45S와 5S rDNA의 염색체상의 위치를 확인한 바, 3쌍의 45S rDNA signal은 4번, 5번 그리고 6번 염색체의 단완 말단 부위에서 관찰되었고, 한 쌍의 5S rDNA signal은 2번 염색체의 동원체 부위에서 관찰되었다.

2배체 담배 Nicotiana plumbaginifolia의 핵형 분석과 rDNAs의 Physical Mapping (Karyotype Analysis and Physical Mapping of rDNAs in Diploid Nicotiana plumbaginifolia)

  • 조혜경;구달회;김수영;방재욱
    • Journal of Plant Biotechnology
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    • 제30권1호
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    • pp.7-11
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    • 2003
  • 2배체 담배인 Nicotiana plumbaginifolia를 대상으로 상염색법과 FISH 기법을 통한 염색체 분석을 수행하여 다음과 같은 결과를 얻었다. N. plumbaginifolia의 체세포 염색체 수는 2n=20이며, arm ratio 비교를 통한 핵형 분석에서 중기 염색체 조성은 3쌍의 중부 염색체 (염색체 1,2 및 7)와 7쌍의 차중부 염색체 (염색체 3, 4, 5, 6, 7, 9 및 10)로 관찰되었다. 염색체의 길이는 2.29~4.50 $\mu\textrm{m}$로 나타났으며, 염색체 1번과 2번은 부수체 염색체로 관찰되었다. 5S와 45S rDNA를 탐침으로 FISH를 수행한 결과 2번 염색체의 동원체 부위에서 한 쌍의 5S signal이 확인되었고, 1번 염색체의 부수체에서 한 쌍의 45S signal이 관찰되었다.

Chromosome Imbalances and Alterations in the p53 Gene in Uterine Myomas from the Same Family Members: Familial Leiomyomatosis in Turkey

  • Hakverdi, Sibel;Demirhan, Osman;Tunc, Erdal;Inandiklioglu, Nihal;Uslu, Inayet Nur;Gungoren, Arif;Erdem, Duygu;Hakverdi, Ali Ulvi
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권2호
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    • pp.651-658
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    • 2013
  • Uterine leiomyomas (UL) are extremely common neoplasms in women of reproductive age, and are associated with a variety of characteristic choromosomal aberrations (CAs). The p53 gene has been reported to play a crucial role in suppressing the growth of a variety of cancer cells. Therefore, the present study investigated the effects of CAs and the p53 gene on ULs. We performed cytogenetic analysis by G-banding in 10 cases undergoing myomectomy or hysterectomy. Fluorescence in situ hybridization (FISH) with a p53 gene probe was also used on interphase nuclei to screen for deletions. In patients, CAs were found in 23.4% of 500 cells analysed, significantly more frequent than in the control group (p<0.001). In the patients, 76% of the abnormalities were structural aberrations (deletions, translocations and breaks), and only 24% were numerical. Deletions were the most common structural aberration observed in CAs. Among these CAs, specific changes in five loci 1q11, 1q42, 2p23, 5q31 and Xp22 have been found in our patients and these changes were not reported previously in UL. The chromosome breaks were more frequent in cases, from high to low, 1, 2, 6, 9, 3, 5, 10 and 12. Chromosome 22, X, 3, 17 and 18 aneuploidy was observed to be the most frequent among all numerical aberrations. We observed a low frequency of p53 losses (2-11%) in our cases. The increased incidence of autosomal deletions, translocations, chromatid breaks and aneuploidy, could contribute to the progression of the disease along with other chromosomal alterations.

Characterization of Interphase Microbial Community in Luzhou-Flavored Liquor Manufacturing Pits of Various Ages by Polyphasic Detection Methods

  • Li, Hui;Huang, Jun;Liu, Xinping;Zhou, Rongqing;Ding, Xiaofei;Xiang, Qianyin;Zhang, Liqiang;Wu, Chongde
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.130-140
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    • 2017
  • It is vital to understand the changing characteristics of interphase microbial communities and interspecies synergism during the fermentation of Chinese liquors. In this study, microbial communities in the three indispensable phases (pit mud, zaopei, and huangshui) of Luzhou-flavored liquor manufacturing pits and their shifts during cellars use were first investigated by polyphasic culture-independent approaches. The archaeal and eubacterial communities in the three phases were quantitatively assessed by combined phospholipid ether lipids/phospholipid fatty acid analysis and fluorescence in situ hybridization. In addition, qualitative information regarding the microbial community was analyzed by PCR-denaturing gradient gel electrophoresis. Results suggested that the interphase microbial community profiles were quite different, and the proportions of specific microbial groups evolved gradually. Anaerobic bacteria and gram-positive bacteria were dominant and their numbers were higher in pit mud ($10^9$ cells/g) than in huangshui ($10^7$ cells/ml) and zaopei ($10^7$ cells/g). Hydrogenotrophic methanogenic archaea were the dominant archaea, and their proportions were virtually unchanged in pit mud (around 65%), whereas they first increased and then decreased in zaopei (59%-82%-47%) and increased with pit age in huangshui (82%-92%). Interactions between microbial communities, especially between eubacteria and methanogens, played a key role in the formation of favorable niches for liquor fermentation. Furthermore, daqu (an essential saccharifying and fermentative agent) and metabolic regulation parameters greatly affected the microbial community.

Cytogenetic Profile of De Novo B lineage Acute Lymphoblastic Leukemia: Determination of Frequency, Distribution Pattern and Identification of Rare and Novel Chromosomal Aberrations in Indian Patients

  • Bhandari, Prerana;Ahmad, Firoz;Dalvi, Rupa;Koppaka, Neeraja;Kokate, Prajakta;Das, Bibhu Ranjan;Mandava, Swarna
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권16호
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    • pp.7219-7229
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    • 2015
  • Background: Chromosomal aberrations identified in acute lymphoblastic leukemia (ALL) have an important role in disease diagnosis, prognosis and management. Information on karyotype and associated clinical parameters are essential to physicians for planning cancer control interventions in different geographical regions. Materials and Methods: In this study, we present the overall frequency and distribution patterns of chromosomal aberrations in both children and adult de novo B lineage ALL Indian patients using conventional cytogenetics, interphase FISH and multiplex RT-PCR. Results: Among the 215 subjects, cytogenetic results were achieved in 172 (80%) patients; normal karyotype represented 37.2% and abnormal 62.8% with a distribution as follows: 15.3% hypodiploidy; 10.3% hyperdiploidy; 15.8% t(9;22); 9.8% t(1;19); 3.7% t(12;21); 2.8% t(4;11); 2.8% complex karyotypes. Apart from these, we observed several novel, rare and common chromosomal rearrangements. Also, FISH studies using LSI extra-signal dual-color probes revealed additional structural or numerical changes. Conclusions: These results demonstrate cytogenetic heterogeneity of ALL and confirm that the incidence of chromosomal abnormalities varies considerably. To the best of our knowledge, this is one of the largest reported series of cytogenetic investigations in Indian B-lineage ALL cases. In addition, ongoing cytogenetic studies are warranted in larger groups of B-lineage ALL cases to identify newly acquired chromosomal abnormalities that may contribute to disease diagnosis and management.

Low Level of TERC Gene Amplification between Chronic Myeloid Leukaemia Patients Resistant and Respond to Imatinib Mesylate Treatment

  • Mohamad Ashari, Zaidatul Shakila;Sulong, Sarina;Hassan, Rosline;Husin, Azlan;Sim, Goh Ai;Wahid, S. Fadilah Abdul
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권4호
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    • pp.1863-1869
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    • 2014
  • The amplification of telomerase component (TERC) gene could play an important role in generation and treatment of haematological malignancies. This present study was aimed to investigate copy number amplification status of TERC gene in chronic myeloid leukaemia (CML) patients who were being treated with imatinib mesylate (IM). Genomic DNA was extracted from peripheral blood of CML-IM Resistant (n=63), CML-IM Respond (n=63) and healthy individuals (n=30). TERC gene copy number predicted (CNP) and copy number calculated (CNC) were determined based on $Taqman^{(R)}$ Copy Number Assay. Fluorescence in situ hybridization (FISH) analysis was performed to confirm the normal signal pattern in C4 (calibrator) for TERC gene. Nine of CML patients showed TERC gene amplification (CNP=3), others had 2 CNP. A total of 17 CML patients expressed CNC>2.31 and the rest had 2.31>CNC>1.5. TERC gene CNP value in healthy individuals was 2 and their CNC value showed in range 1.59-2.31. The average CNC TERC gene copy number was 2.07, 1.99 and 1.94 in CML-IM Resistant patients, CML-IM Respond and healthy groups, respectively. No significant difference of TERC gene amplification observed between CML-IM Resistant and CML-IM Respond patients. Low levels of TERC gene amplification might not have a huge impact in haematological disorders especially in terms of resistance towards IM treatment.