• Title/Summary/Keyword: Chromosome analysis

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Construction and analysis of painting probe for homogeneously staining regions in human neuroblastoma cell line IMR-32

  • Park, Sun-Hwa;Kim, Ho-Chung;Chun, Yong-Hyuck
    • Journal of Genetic Medicine
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    • v.1 no.1
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    • pp.45-50
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    • 1997
  • Neuroblastoma, a pediatric malignant neoplasm of neural crest origin, has a wide range of clinical virulence. The mechanisms contributing to the development of neuroblastomas are largely unclear, but non-random chromosomal changes identified over the past years suggest the involvement of genetic alterations. Amplification of the human N-myc proto-oncogene is frequently seen either in extrachromosomal double minutes or in homogeneously staining regions (HSRs) of aggressively growing neuroblastomas. N-myc maps to chromosome 2 band 24, but HSR have never been observed at this band, suggesting transposition of N-myc during amplification. We have constructed and analyzed the region-specific painting probe for HSR in neuroblastoma IMR-32 to determine the derivative chromosomes. Microdissection was performed on HSR using an inverted microscope with the help of microglass needles and an micromanipulator. We pretreated the microdissected fragments with Topoisomerase I which catalyzes the relaxation of supercolled DNA, and performed two initial rounds of DNA synthesis with T7 DNA polymerase followed by conventional PCR to enable the reliable preparation of Fluorescent in situ hybridization probe from a single microdissected chromosome. With this method, it was possible to construct the region-specific painting probe for HSR. The probe hybridized specifically to the HSRs of IMR-32, and to 2p24, 2p13 of normal chromosome. Our results suggest there was coamplification of N-myc together with DNA of the chromosome 2p24 and 2p13. Moreover, the fluorescent signals for the amplified chromosomal regions in IMR-32 cells were also easily recognized at a Thus this painting probe can be applied to detect the similar amplification of N-myc in neuroblastoma tissue, and the probe pool for HSR may be used to identify the cancer-relevant genes.

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Prenatal diagnosis of interchromosomal insertion of Y chromosome heterochromatin in a family

  • Lee, Bom-Yi;Park, Ju-Yeon;Lee, Yeon-Woo;Oh, Ah-Rum;Lee, Shin-Young;Park, So-Yeon;Ryu, Hyun-Mee;Lee, Si-Won
    • Journal of Genetic Medicine
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    • v.14 no.2
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    • pp.62-66
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    • 2017
  • Interchromosomal insertion of Y chromosome heterochromatin in an autosome was identified in a fetus and a family. A fetal karyotype was analyzed as 46,XX,dup(7)(?q22q21.1) in a referred amniocentesis at 16 weeks of gestation for advanced maternal age. In the familial karyotype analyses for identification of der(7), the mother, the first daughter and the maternal grandmother showed the same der(7) as the fetus's. CBG-banding was positive at 7q22 region of der(7) that indicated inserted material was originated from heterochromatin. The origin of heterochromatic insertion region in der(7) of the fetus and the mother was found in Yq12 region by fluorescent in situ hybridization with a DYZ1 probe. In the specific analysis of Y chromosomal heterochromatic region of ins(7;Y) of the mother, 15 sequence tagged sites from Yp11.3 region including SRY to Yq11.223 region was not detected. Final karyotypes of the mother, the first daughter and the maternal grandmother were reported as 46,XX,der(7)ins(7;Y)(q21.3;q12q12). All female carriers of ins(7;Y) in the family showed normal phenotype and the mother and the maternal grandmother were fertile. A healthy girl was born at term. We report a rare case of familial interchromosomal insertion of Y chromosome heterochromatin detected only in female family members with normal phenotype that was diagnosed prenatally.

The Effect of 5-FU and Radiation on A549 Cells In Vitro (시험관내의 인체폐암 세포 A549의 방사선 및 5-Fluorouracil에 대한 효과)

  • Lee, Myung-Za;Chun, Ha-Chong;Lee, Won-Young
    • Radiation Oncology Journal
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    • v.7 no.1
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    • pp.1-13
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    • 1989
  • Effects of ionizing radiation alone and combined with chemotherapy on tumor growth and it's clonal specificity monitored by changes in distribution of chromosome number were studies in A549 ceil line originated from human adenocarcinoma of the lung. Radiation (300 rad, 600 rad and 900 rad) were delivered with or without 5-FU. Forty eight hours later, 57.5% of growth inhibition of cell w8s seen in cells treated with 5-FU concentration of $0.4{\mu}g/ml$ for 24hr exposure. Cell survival curves after radiation with and without 5-FU were made. Chromosomal analysis of cells in metaphase in control, and in cells treated with 300 rad of radiation, or $0.4{\mu}g/ml$ of 5-FU treatment, and combined treatment of both were done to examine the changes in ploidy and number of chromosome. Radiation combined with S-FU enhanced growth inhibition of A549 cells. However, no evidence of synergegic effects in growth. inhibition was observed in the cells treated with the combination therapy. Pattern of chromosomal distribution of survived cells were shifted from hyperploidy to hypoploidy by single dose of radiation (300 rad). As radiation dose increased a large number of hypoploidy cells were observed. Following treatment of cells with 5-FU, chomosomal distribution of survived cells were also shifted to hypodiploidy which were seen in cells treated with radiation, The ceil treated with 5-FU and fellowed by radiation within 24 hrs had cell with increased number of hypodiploidy cells. Almost same type of chromosomal changes were reproduced in cells treated with combined treatment with radiation and 5-FU. Minor differences were that cells with fewer number of chromosome were more frequent in cells treated with combined therapy. Further increase in cells of hypoploidy (93%) having 1-10 chromosome were induced by additional radiation. Therefore, the enhanced therapeutic effect of 5-FU combined with radiation of A549 cells appeared to be additive rather than synergistic.

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Complete genome sequence of Salmonella Enteritidis MFDS1004839 isolated from food (식품에서 분리된 Salmonella Enteritidis MFDS1004839의 유전체 서열 분석)

  • Lee, Woojung;Park, Sewook;Yoo, Ran Hee;Joo, In-Sun;Kwak, Hyo Sun;Kim, Soon Han
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.164-166
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    • 2018
  • Salmonella enterica subsp. enterica is a foodborne pathogen that has been detected throughout the world. Here, we present the complete genome sequence of Salmonella Enteritidis isolated from a commercial kimbap that caused foodborne illness in the Republic of Korea in 2014. Complete genome sequence analysis of Salmonella Enteritidis MFDS1004839 revealed a 4,679,649 bp chromosome and a 96,994 bp plasmid, with G + C contents of 52.2% and 49.3%, respectively. The chromosome and plasmid genome included 4,482 predicted protein-coding sequences, 84 tRNAs and 22 rRNAs genes.

Chromosomal study of the lenoks, Brachymystax (Salmoniformes, Salmonidae) from the South of the Russian Far East

  • Kartavtseva, I.V.;Ginatulina, L.K.;Nemkova, G.A.;Shedko, S.V.
    • Journal of Species Research
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    • v.2 no.1
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    • pp.91-98
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    • 2013
  • An investigation of the karyotypes of two species of the genus Brachymystax (B. lenok and B. tumensis) has been done for the Russia Primorye rivers running to the East Sea basin, and others belonging to Amur basin. Based on the analysis of two species chromosome characteristics, combined with original and literary data, four cytotypes have been described. One of these cytotypes (Cytotype I: 2n=90, NF=110-118) was the most common. This common cytotype belongs to B. tumensis from the rivers of the East Sea basin and B. lenok from the rivers of the Amur basin, i.e. extends to the zones of allopatry. In the rivers of the Amur river basin, in the zone of the sympatric habitat of two species, each taxon has karyotypes with different chromosome numbers, B. tumensis (2n=92) and B. lenok (2n=90). Because of the ability to determine a number of the chromosome arms for these two species, additional cytotype have been identified for B. tumensis: Cytotype II with 2n=92, NF=110-124 in the rivers basins of the Yellow sea and Amur river and for B. lenok three cytotypes: Cytotype I: 2n=90, NF=110 in the Amur river basin; Cytotype III with 2n=90, NF=106-126 in the Amur river basin and Cytotypes IV with 2n=92, NF=102 in the Baikal lake.

Karyotype Analysis and rDNA Physical Mapping in Rye (Secale cereale L.) (호밀(Secale cereale L.)의 핵형분석과 rDNA의 Physical Mapping)

  • Lee, Joon Soo;Seo, Bong Bo;Kim, Min
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.163-168
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    • 2010
  • This study was carried out to determine the chromosomal localization of the 5S and 18S-26S ribosomal DNA(rDNA) genes by means of fluorescence in situ hybridization(FISH) techniques, and the constitutive heterochromatin detected by means of Gimsa C-banding technique in rye(Secale cereale L.). The somatic chromosomes number was 2n=14. The karyotype consists of four pairs of metacentrics(chromosomes 1, 2, 3, and 7) and three pairs of submetacentrics(chromosomes 4, 5, and 6). Secondary constrictions appeared in the short arm of chromosome 1. The 5S rDNA genes have been located on two pairs of chromosomes 1 and 5, and 18S-26S rDNAs genes have been located on one pair of chromosome 1. 5S rDNA genes were detected on the distal region of the secondary constrictions in nucleolus organizer regions(NOR) in chromosome 1, and other detected on the intercalary region in the short arm of chromosome 5.

Assessment of genotoxicity of Ssanghwa-tang, an herbal formula, by using bacterial reverse mutation, chromosome aberration, and in vivo micronucleus tests

  • Jang, Ji-Hye;Seo, Chang-Seob;Lee, Mee-Young;Shin, Hyeun-Kyoo;Han, Su-Cheol;Ha, Hyekyung
    • The Journal of Korean Medicine
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    • v.42 no.4
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    • pp.25-39
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    • 2021
  • Objectives: Ssanghwa-tang (SHT) is a traditional herbal formula comprising nine medicinal herbs, and it is used for reducing fatigue in Korea. SHT exerts various effects such as anti-inflammatory, antioxidant, and anti-aging activities, and protection against acute hepatotoxicity. However, the genotoxicity of SHT has not yet been established. Methods: Ten components were identified in SHT water extract by using high-performance liquid chromatography analysis. We assessed the genotoxicity of SHT by using bacterial reverse mutation (Ames test), chromosome aberration, and in vivo micronucleus tests. Results: The contents of paeoniflorin, glycyrrhizin, and liquiritin apioside in SHT were 15.57, 6.94, and 3.48 mg/g extract, respectively. SHT did not increase the revertant colonies of Salmonella typhimurium and Escherichia coli strains in the presence or absence of metabolic activity. Although SHT did not induce structurally abnormal chromosomes in Chinese hamster lung (CHL) cells in the presence of metabolic activity, the number of structurally aberrated chromosomes increased dose-dependently in the absence of metabolic activity. In the in vivo micronucleus test, SHT did not affect the formation of micronuclei compared with the vehicle control. Conclusions: Genotoxicity of SHT was not observed in the Ames test and in vivo micronucleus test. However, based on the results of chromosome aberration test, it can be presumed that SHT has the potential to induce genotoxicity because it induced structurally abnormal chromosomes in the absence of metabolic activity.

Genome-wide SNP analysis provides insights into the XX/XY sex-determination system in silver barb (Barbonymus gonionotus)

  • Visarut Chailertrit;Thitipong Panthum;Lalida Kongkaew;Piangjai Chalermwong;Worapong Singchat;Syed Farhan Ahmad;Ekaphan Kraichak;Narongrit Muangmai;Prateep Duengkae;Surin Peyachoknagul;Kyudong Han;Kornsorn Srikulnath
    • Genomics & Informatics
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    • v.21 no.4
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    • pp.47.1-47.12
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    • 2023
  • Silver barb (Barbonymus gonionotus) is among the most economically important freshwater fish species in Thailand. It ranks fourth in economic value and third in production weight for fisheries and culture in Thailand. An XX/XY sex-determination system based on gynogenesis was previously reported for this fish. In this study, the molecular basis underlying the sex-determination system was further investigated. Genome-wide single-nucleotide polymorphism data were generated for 32 captive-bred silver barb individuals, previously scored by phenotypic sex, to identify sex-linked regions associated with sex determination. Sixty-three male-linked loci, indicating putative XY chromosomes, were identified. Male-specific loci were not observed, which indicates that the putative Y chromosome is young and the sex determination region is cryptic. A homology search revealed that most male-linked loci were homologous to the Mariner/Tc1 and Gypsy transposable elements and are probably the remnants of an initial accumulation of repeats on the Y chromosome from the early stages of sex chromosome differentiation. This research provides convincing insights into the mechanism of sex determination and reveals the potential sex determination regions in silver barb. The study provides the basic data necessary for increasing the commercial value of silver barbs through genetic improvements.

Pseudoisodicentric X chromosome in a female with primary amenorrhea (원발성 무월경 여성에서 관찰된 Pseudoisodicentric X 염색체)

  • Park, Sang-Hee;Shim, Sung-Han;Chin, Mi-Uk;Kang, Su-Jin;Bae, Sung-Mi;Sohn, Soo-Min;Cha, Dong-Hyun;Yoon, Tae-Ki;Cho, Jung-Hyun
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.61-64
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    • 2008
  • A 24-year-old female with primary amenorrhea was referred for a chromosome study. The karyotype of the patient was 46,X,der(X) under initial GTG-banding analysis. Fluorescence in situ hybridization (FISH) analysis with an LSI Kallmann (KAL) region probe [probes for Xp22.3(KAL) and CEP(X) for control] was carried out. The abnormal chromosome was KAL- and CEP(X)${\times}2$. In addition, interphase FISH analysis revealed the patient to be mosaic for two different cell lines: 90% of cells had three signals and 10% of the cells had only one signal for CEP(X). Based on these results, the karyotype of the patient was 45,X/46,X,psu idic(X)(p22.1), which is partial trisomy for Xqter${\rightarrow}$Xp22.1 and partial monosomy for Xpter${\rightarrow}$Xp22.1. This karyotype was considered a variant of Turner syndrome. In summary, Idic(X) and low-level mosaicism was successfully characterized by FISH analysis with a CEP(X) probe.

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QTL Analysis of Concerned on Ideal Plant Form in Rice (벼의 이상적인 초형에 관여하는 QTL 분석)

  • Chung, Il Kyung;Kim, Kyung-Min
    • Korean Journal of Plant Resources
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    • v.30 no.2
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    • pp.213-218
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    • 2017
  • Rice is the staple food of at least half of the world's population. Due to global warming, the weather is difficult to forecast nowadays. Therefore, it is necessary to breed various breeding to respond to such changes in the environment. This study was conducted to analyze the QTL about plant form, culm length, ear number and ear length by using 120 lines by anther culture, a cross between the Indica variety Cheongcheong and Japonica variety Nagdong. DNA marker was selected on the QTLs gene, and the following results were obtained. CNDH (Cheongcheong Nagdong Doubled Haploid) lines frequency distribution table curves about culm length, ear number and ear length exhibited showed a continuous variation close to a normal distribution. QTL analysis result, on culm length qPlL1-1 and qPlL1-2 were detected on the chromosome 1 and qPlL5 was detected on the chromosome 5. However, on ear length qPL2, qPL3 and qPL10, were detected on the chromosome 2, 3 and 10, while on ear number qPN1-1 and qPN1-2 were detected on the chromosome 1, qPN9 was detected on the chromosome 9. The QTLs related to culm length was found to chromosomes 5 and LOD scores were 3.81. The QTLs related to ear length was found to chromosomes 2 and 3 LOD scores were 7.13 and 3.20. The QTLs related to ear number was found to chromosome 9 and LOD scores were 4.27. Twenty two (22) Japonica cultivars and 12 Indica cultivars were analyzed polymorphisms, using selected 9 markers from the result about plant form analysis. RM5311, RM555 and RM8111 about the culm length, the ear length and number of ear were selected on the standard of Cheongcheong and Nagdong. Each rate of concordances about the culm length, the ear length and number of ear are 44.11%, 41.17% and 44.11%.