• Title/Summary/Keyword: chromosomal DNA

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Cytogenetic Analysis in Korean Head and Neck Cancer Cell Lines: Comparative Genomic Hybridization(CGH) and Array-CGH (두경부 편평상피세포암 세포주의 염색체 이상 분석: 비교유전체보합법과 Array 비교유전체보합법)

  • Shin, You-Ree;Park, Soo-Yeun;Lee, Dong-Wook;Kim, Han-Su;Go, Young-Min;Park, Hyun-Joo;Choung, Sung-Min
    • Korean Journal of Head & Neck Oncology
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    • v.24 no.1
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    • pp.33-42
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    • 2008
  • Head and neck squamous cell carcinoma(HNSCC) is notorious for its poor outcome and increasing incidence. But, the studies of cytogenetic analysis in HNSCC are relatively rare, because of difficulties in culturing solid tumor cells and complexity in chromosomal DNA abberations associated with the lesions. The purpose of this study is to evaluate the location of chromosomal aberrations in Korean HNSCC cell lines (SNU-1041, 1066, and 1076) with comparative genomic hybridization(CGH) and array based CGH(array-CGH). Chromosomal gains of 3q23-q27, 5p13-p15.3, 7p21-pter, 8q11.2-q12, 8q21.1-qter, 9q22-q34, 16q22-q24, and 20q11.2-qter, as well as chromosomal losses on 3p10-p14 were found in all 3 SNU cell lines. Losses on 3p15- p23, 4q22-q27, 4q31.3-qter, 6q14-q15, 7q31-q34, 8p12-pter, 18q21-q23, and 21q11.2-q12 were observed in 2 of 3 cell lines. In array-CGH, many genes were altered including gains of PIK3CA, MYC, EVI1, MAD1L1 genes and losses of SERPIN genes. These aberrations of gene and chromosome coincide with other results of study, generally. These data about the patterns of chromosomal aberrations could be a basic step for understanding more detailed genetic events in the carcinogenesis and also provide information for diagosis and treatment in HNSCC.

Clinical, Cytogenetic and CYP1A1 exon-1 Gene Mutation Analysis of Beedi Workers in Vellore Region, Tamil Nadu

  • Sundaramoorthy, Rajiv;Srinivasan, Vasanth;Gujar, Jidnyasa;Sen, Ayantika;Sekar, Nishu;Abilash, Valsala Gopalakrishnan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7555-7560
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    • 2013
  • Background: Beedi rollers are exposed to unburnt tobacco dust through cutaneous and pharyngeal route and it is extremely harmful to the body since it is carcinogenic in nature and can cause cancer during long exposure. This indicates that occupational exposure to tobacco imposes considerable genotoxicity among beedi workers. Materials and Methods: In the present study, 27 beedi workers and age and sex matched controls were enrolled for clinical, cytogenetics and molecular analysis. Clinical features were recorded. The workers were in the age group of 28-67 years and were workers exposure from 8-60 years. Blood samples were collected from workers and control subjects and lymphocyte cultures were carried out by using standard technique, slides were prepared and 50 metaphases were scored for each sample to find the chromosomal abnormalities. For molecular analysis the genomic DNA was extracted from peripheral blood, to screen the variations in gene, the exon 1 of CYP1A1 gene was amplified by polymerase chain reaction (PCR) and then screened with Single Strand Conformation Polymorphism (SSCP) analysis. Results: A statistically significant increase was observed in the frequencies of chromosomal aberrations in exposed groups when compared to the respective controls and variations observed in Exon 1 of CYP1A1(Cytochrome P450, family 1, subfamily A, polypeptide 1) gene. Conclusions: This study shows that, the toxicants present in the beedi that enter into human body causes disturbance to normal state and behavior of the chromosomes which results in reshuffling of hereditary material causing chromosomal aberrations and genomic variations.

Genotoxicity Assessment of Gardenia Yellow using Short-term Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Ahn, Jun-Ho;Choi, Seon-A;Noh, Hong-June;Seo, Young-R.;Oh, Se-Wook;Lee, Michael
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.257-264
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    • 2009
  • Gardenia yellow, extracted from gardenia fruit, has been widely used as a coloring agent for foods, and thus, safety of its usage is of prime importance. In the current study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of gardenia yellow. The gardenia yellow used was found to contain 0.057 mg/g of genipin, a known biologically active compound of the gardenia fruit extract. Ames test did not reveal any positive results. No clastogenicity was detected by a chromosomal aberration test, even on evaluation at the highest feasible concentration of gardenia yellow. Gardenia yellow was also shown to be non-genotoxic using an in vitro comet assay and a micronucleus test with L5178Y cells, although a marginal increase in DNA damage and micronuclei frequency was reported in the respective assays. Additionally, in vivo micronucleus test results clearly demonstrated that oral administration of gardenia yellow did not induce micronuclei formation in the bone marrow cells of male ICR mice. Taken together, our results indicate that gardenia yellow is not mutagenic to bacterial cells, and that it does not cause chromosomal damage in mammalian cells, either in vitro or in vivo.

Molecular Cloning, Chromosomal Integration and Expression of the Homoserine Kinase gene THR1 of Saccharomyces cerevisiae (트레오닌 생합성에 관여하는 효모유전자 THR1의 클로님, 염색체통합 및 발현)

  • 최명숙;이호주
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.16-24
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    • 1991
  • The yeast gene THR1 encodes the homoserine kinase (EC 2.7.1.39: HKase) which catalyses the first step of the threonine specific arm at the end of the common pathway for methionine and threonine biosynthesis. A recombinant plasmid pMC3 (12.6 kilobase pairs, vector YCp50) has been cloned into E. coli HB101 from a yeast genomic library through its complementing activity of a thr1 mutation in a yeast recipient strain M39-1D. When subcloned into pMC32 (8.6kbp, vector YRp7) and pMC35 (8.3 kbp, vector YIp5), the HindIII fragment (2.7 kbp) of pMC3 insery was positive in the thrI complementing activity in both yeast and E. coli auxotrophic strains. The linearized pMC35 was introduced into the original recipient yeast strain and the mitotically stable chromosomal integrant was identified among the transformants. Through the tetrad analysis, the integration site of the pMC35 was localized to the region of THR1 structural gene at an expected genetic distance of approximately 11.1 cM from the ARG4 locus on the right arm of the yeast chromosome VIII. When episomically introduced into the auxotrophic cells and cultured in Thr omission liquid medium, the cloned gene overexpressed the HKase in the order of thirteen to fifteenfold, as compared with a wildtype. HKase levels are repressed by addition of threonine at the amount of 300 mg/l and 1, 190 mg/l for pMC32 and pMC3, respectively. Data from genetic analysis and HKase response thus support that the cloned HindIII yeast DNA fragment contains the yeast thr1 structural gene, along with necessary regulatory components for control of its proper expression.

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In Vitro Genotoxicity Assessment of a Novel Resveratrol Analogue, HS-1793

  • Jeong, Min Ho;Yang, Kwangmo;Lee, Chang Geun;Jeong, Dong Hyeok;Park, You Soo;Choi, Yoo Jin;Kim, Joong Sun;Oh, Su Jung;Jeong, Soo Kyung;Jo, Wol Soon
    • Toxicological Research
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    • v.30 no.3
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    • pp.211-220
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    • 2014
  • Resveratrol has received considerable attention as a polyphenol with various biological effects such as anti-inflammatory, anti-oxidant, anti-mutagenic, anti-carcinogenic, and cardioprotective properties. As part of the overall safety assessment of HS-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, we assessed genotoxicity in three in vitro assays: a bacterial mutation assay, a comet assay, and a chromosomal aberration assay. In the bacterial reverse mutation assay, HS-1793 did not increase revertant colony numbers in S. typhimurium strains (TA98, TA100, TA1535 and TA1537) or an E. coli strain (WP2 uvrA) regardless of metabolic activation. HS-1793 showed no evidence of genotoxic activity such as DNA damage on L5178Y $Tk^{+/-}$ mouse lymphoma cells with or without the S9 mix in the in vitro comet assay. No statistically significant differences in the incidence of chromosomal aberrations following HS-1793 treatment was observed on Chinese hamster lung cells exposed with or without the S9 mix. These results provide additional evidence that HS-1793 is non-genotoxic at the dose tested in three standard tests and further supports the generally recognized as safe determination of HS-1793 during early drug development.

Site-speci fic Inactivation o meso-Diaminopimelate-dehydrogenase Gene (ddh) in a Lysine-producing Brevibacterium lactofementum. (Brevibacterium lactofermentum 에서 meso-Diaminopimelate-dehydrogenase Gene (ddh)의 Site-specific Inactivation)

  • 김옥미;박선희;이갑랑
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.387-392
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    • 1998
  • Brevibacterium lactofermentum, a gram-positive bacteria, has both the diaminopimelate (DAP) pathway and meso-DAP-dehydrogenase (DDH) pathway for L-lysine biosynthesis. To investigate importance of DDH pathway and the related ddh gene in lysine production, we introduced site-specific mutagenesis technique. A 300 bp DNA fragment central to the meso-DAP-dehydrogenase gene (ddh) of B. lactofermentum was used to inactive chromosomal ddh gene via homologous recombination. Southern hybridization analysis confirmed that the chromosomal ddh gene was disrupted by the vector sequence. The B. lactofementum ddh mutant obtained have an inactive DDH pathway. The results reveal that inactivation of the ddh gene in B. lactofermentum leads to dramatic reduction of lysine production as well as decrease of the growth rate, indicating that the DDH pathway is essential for high-level lysine production as well as biosynthesis of meso-DAP.

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Age-related Increase of Sister Chromatid Exchange Frequency in Bone Marrow Cells of Senescence Accelerated Mouse and Its Inhibition by Chronic Treatment of Ginseng

  • Lim, Heung-Bin;Sohn, Hyung-Ok;Lee, Young-Gu;Kim, Seung-Hyung;Lee, Dong-Wook
    • Toxicological Research
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    • v.11 no.2
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    • pp.261-266
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    • 1995
  • Age-related change in the frequency of spontaneous sister chromatid exchange (SCE) and chromosornal aberrations were investigated in bone marrow cells of accelerated senescence-resistant mice (SAM R1) and senescence accelerated ones (SAM P1). And the effect of chronic treatment of ginseng extract (Panax ginseng C.A. Meyer) on these chromosomal abnormalities was tested in SAM P1. SCE frequency in the cells was progressively increased with age in both mice, but it was consistently higher in SAM P1 than in SAM R1 at all corresponding age. Chromosomal aberrations were, however, not significantly changed with age except that it was slightly increased in only aged SAM P1. Interestingly, the rate of these genetic instabilities in SAM P1 was remarkably retarded by long-term administration of ginseng water extract (0.05% in drinking water). These results suggest that frequency of spontaneous SCE in bone marrow cells increase in parallel with senescence of the mice, and SAM P1 is in the condition of being more exposed than SAM R1 to DNA damaging factors. These also indicate that long-term treatment of ginseng may reduce the genetic damage.

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Agouti Gene의 Human Homologue의 Molecular Structure와 Chromosomal Mapping

  • Heajoon Y. Kwon;Scott J. Bultman;Christiane Loffler;Chen, Wen-Ji;Paul J. Furdon;John G. Powell;Usala, Anton-Lewis;William Wilkison;Ingo Hansman
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.55-64
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    • 1996
  • mouse chromesome2에 있는 agouti locus는 정상적으로는 털색깔을 조절하는 gene이다. mouse agouti gene은 최근에 cloning 되었고 131 amino acid peptide와 consensus signal peptide를 encode한다고 보고되었다. 이 논문에서 interspecies-DNA hybridization approach를 이용하여 mouse agouti gene의 human homologue를 cloning 하였다. Sequence analysis 결과, 이는 mouse gene에 85% 유사하였고 consensus signal peptide sequence 를 포함하는 132 amino acid를 coding하였다. somatic-cell hybrid mapping pannel과 Fluorescence-in-situ hybridization에 의한 chromosomal mapping을 한 결과, agouti gene은 MODY (maturity onset diabetes of the young), myeloid leukemia locus 등이 위치한 human chromosome 20q 11.2에 mapping 되었다. 성인 tissue로부터 추출한 RNA를 이용한 발현연구에 의하면 human agouti gene은 adipose tissue와 teatis에 발현되었다.

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PCR-Based Polymorphic Analysis for the Y Chromosomal Loci DYS19 and DXYS5Y (47z) in the Korean Population

  • Shin, Dong-Jik;Kim, Yung-Jin;Kim, Wook
    • Animal cells and systems
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    • v.2 no.2
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    • pp.281-285
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    • 1998
  • We examined Y chromosomal DNA polymorphisms at the DYS19 and DXYS5Y loci in a total of 480 unrelated male samples from the Korean population. All five common alleles were identified at the tetranucleotide microsatellite locus DYS19 in this study. The C allele was the most frequent (212/480), followed by D (136/480), B (75/480), E (36/480) and A (21/480) allele. The frequency of Y2 allele at the DXYS5Y locus was found to be 4.6% (22/480). Combining the allelic variation at these two loci resulted in a total of 9 combination haplotypes. The mean combination haplotype diversity wIns 0.72. Based on the results of these two loci, Korean and Japanese populations may share some common genetic structure that is rare or absent in the other ethnic groups. The genetic similarity between Korean and Japanese populations may be due to the large infusion of Y chromosomes through the Yayoi migration starting 2,300 years ago from Korea to Japan.

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Cloning and Expression of the Structural Gene for Alcohol Dehydrogenase of Zymomonas mobilis in Escherichia coli (Zymomonas mobilis 알코올 탈수소 효소 유전자의 Cloning과 Escherichia coli 에서의 발현)

  • Yoon, Ki-Hong;Shin, Byung-Sik;M.Y Pack
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.301-306
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    • 1989
  • A genomic library of Zymomonas mobilis DNA was constructed in Escherichia coli using plasmid pUC9 Allyl alcohol was used to screen a genomic clone expressing alcohol dehydrogenase. The plasmids isolated from two clones, which were sensitive to allyl alcohol, were found to be related and to share a common 2.6 kb fragment encoding alcohol dehydrogenase II identified as one of two isozymes in Z. mobilis by staining for alcohol dehydrogenase activity on polyacrylamide gel and spectrophotometric analysis of several substrate oxidations.

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