• Title/Summary/Keyword: Chromosomal

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Chromosomal Localization and Distribution of the Telomeric DNA in Cattle and Pigs (소, 돼지 염색체의 telomeric DNA 분포 양상)

  • Sohn, S.H.;Multani, A.S.;Pathak, S.;Cho, E.J.;Ha, H.B.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.547-554
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    • 2004
  • Telomeres are nucleoprotein structures at the ends of chromosomes consisting of tandem repeat sequences of . (TTAGGG)n. Telomeres serve as guardians of the genome, protect individual chromosomes within the nucleus, and help in meiotic pairing of homologous chromosomes. To investigate the telomere distributions of cattle and pig chromosomes, fluorescence in situ hybridization(FISH) was carried out on metaphase spreads of in vitro fibroblast cultures from Holstein and Landrace using a human telomeric DNA repeat probe. Results indicate that the distinct double spots on both ends of chromosomes of cattle and pigs were observed. In cattle, there was a random variation in the intensity of telomere signals among chromosomes. In pigs, an interstitial telomeric signal was observed on the chromosome 6q1 of all the cells examined. According to quantitative fluorescence in situ hybridization(Q-FISH) analysis, some chromosomes had consistently much more telorneres at one end of chromosomes. In general, both species had consistently much more telomeres at q-end than p-end on most of chromosomes. The relative amount of telomeres on bovine chromosomes was higher than that on pig chromosomes. In additions, Y chromosome had the highest relative amount of telorneres in cattle and pigs.

Cytoplasmic Inheritance of High Tillering and Earing Characters of a Korean Local Maize Line(MET) (다수.다얼성 옥수수(NET)의 세포질적 유전)

  • Han, C.H.;Lee, I.S.;Choe, B.H.;Park, K.Y.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.1
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    • pp.55-61
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    • 1984
  • A Korean local maize line, MET, which has multi-ears and tillers has been proved as a potential source for silage production. However, no fundamental genetic nature for the line has been investigated. Therefore, this study was done to find genetic information on the multi-earing and -tillering habits of MET line. MET line and a hybrid. (Mo 17 ${\times}$ B68), with monoculm and single ear per plant were used for production of F$_1$(F$\_$1-12/ and F$\_$1-21/), F$\_$2-12/, F$\_$2-21/, BC$\_$1-12/ and BC$\_$1-21/ generations. From the comparison of reciprocal crosses, it was found that the tillering and earing habits of the MET line are controlled by cytoplasmic factors. The tiller and ear numbers, and barren ears were all characters associated with the MET cytoplasm. The cytoplasmic effect of MET on tiller and ear numb en was not evident in F$_1$ generation, probably because of suppressing effect of heterosis on appearance of tillers or ears. Genetic parameters for the gene action for both tiller and ear number also indicated a lack of mono- or digenic-chromosomal gene effects. The heritability (broad) was very low for both characters. Therefore, it is strongly concluded that the tillering and earing characters of MET line are due to cytoplasmic reasons.

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Immunofluorescence and Electron Microscopic Study on the Artificial Insemination and Rotation-Shift Behaviors of the Bipolar Spindle Fiber in U. unicinctus Egg (U. unicinctus 난자의 인공수정과 감수분열 장치의 회전-이동행위에 관한 면역형광현미경 및 전자현미경적 연구)

  • Kwon, Hyuk-Jae;Jeong, Jin-Wook;Kim, Wan-Jong;Shin, Kil-Sang
    • Applied Microscopy
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    • v.33 no.2
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    • pp.105-116
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    • 2003
  • In Vitro fertilization of U. unicinctus eggs observed by immunofluorescence and electron microscopes revealed an overview of the meiotic pattern of the tide animals. The eggs have been fertilized early at germinal vesicle stage, followed by germinal vesicle break down (GVBD), but pre-mitotic aster like structure could not be resolved by the methods employed in this work. The meiotic features, such as rotation-shift movement of spindle fibers, behavior of spermatozoonmonaster in the egg cytoplasm and active spindle fiber of the 1st polar body, have been observed. The antitubulin-FITC fluorescence show the 2nd meiotic apparatus appeared firstly parallel to the tangential line of the oolemma, proceeding the meiosis, its bipolarity is rotated and shifted towards the oolemma. The polar bodysite of the oolemma was not amorphous, but active in a sense of anti-tubulin-FITC reactions during the extrusions of the polar bodies. The immunofluorescence reactions of the spermatozoon centriole appeared at a later stage of the 2nd meiosis. During the time periods, the fertilized spermatozoon resided in the egg cytoplasm. Activating the centrioles, spermatozoon approaches towards the chromosomal materials of the 2nd oocyte. This suggests that spermatozoon centrioles initiate and play a roll to fuse male and female pronuclei.

Gene Cloning, Nucleotide Sequence and Efficent Expression of Peptidyl proryl cis-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus의 Peptidyl Prolyl cis-trans Isomerase 유전자 분리 염기배열 및 발현)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.452-458
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    • 1996
  • A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

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Genetic Analysis on Floury Endosperm Characteristics of 'Namil(SA)-flo1', a Japonica Rice Mutant Line (남일벼 돌연변이 후대 계통 'Namil(SA)-flo1'의 분질배유 특성에 대한 유전분석)

  • Mo, Young-Jun;Jeung, Ji-Ung;Kang, Kyung-Ho;Lee, Jeom-Sig;Kim, Bo-Kyeong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.3
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    • pp.283-291
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    • 2013
  • Rice varieties with suitable flour-making quality are required to promote rice processed-food industry and boost rice consumption in Korea. 'Namil (SA)-flo1' is an advanced mutant line with floury endosperm which shows good flour-making quality under dry-milling process. Genetic analysis was carried out to localize the chromosomal region responsible for the floury endosperm of 'Namil (SA)- flo1'. By using 94 F2 progenies, which were derived from 'Namil (SA)-flo1' ${\times}$ 'Milyang 23', floury grains percentage was investigated as phenotypic data, and genotyping was conducted with 54 SSR markers. Association analysis showed that the target genetic region for floury endosperm is on middle-low region of chromosome 5. Through further association analysis with increased number of SSR markers on chromosome 5, we found that genotypic variation in RM164 explains 79.7% of the variation in floury grains percentage of F2:3 seeds. The floury endosperm locus was localized on 17.7-20.7 Mbp region of chromosome 5 and will be further analyzed for fine mapping and gene identification.

Evaluation of DNA Damage and Repair Kinetics in the Earthworm (Eisenia fetida) Exposed to Radiation and Mercury (방사선과 수은에 의해 유도된 Eisenia fetida 체강세포의 DNA 손상 및 수복 평가)

  • Ryu, Tae-Ho;Nili, Mohammad;An, Kwang-Guk;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.68-73
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    • 2011
  • The single cell gel electrophoresis (SCGE) assay is a microelectrophoretic technique for assessments of DNA damage at the level of the individual eukaryotic cell. The SCGE assay, due to its simplicity, sensitivity and need of a few cells, has advantages compared to other genomic damage assays such as sister chromatid exchange, chromosomal aberration and micronucleus test. In this study, investigated were the levels of DNA damage and the repair kinetics in the coelomocytes of Eisenia fetida treated with HgCl2 and ionizing radiation by means of the SCGE assay. For detecting DNA damage and repair in coelomocytes, earthworms (E. fetida) were irradiated with six doses of ${\gamma}$-rays (0, 2.5, 5, 10, 20 and 50 Gy) and in vivo exposed to mercuric chloride at 0, 80 and 160 mg $kg^{-1}$ for 48 hours. Then the Olive tail moments were measured during 0~12 hours after irradiation and 0~72 hours after Hg treatment. The results showed that the more the oxidative stress was induced by mercury and radiation, the longer the repair time was required. Also, the results suggest that the SCGE assay may be used as an important tool for comparison of the sensitivity of different species to oxidative stresses.

Restriction Fragment Length Polymorphism of Mycobacterium Tuberculosis in a University Hospital in Seoul (서울의 한 대학병원에서 동정된 결핵균 균주의 RFLP 양상)

  • Kim, Woo-Jin;Yim, Jae-Joon;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.308-314
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    • 2000
  • Background : DNA fingerprinting of Mycobacterium tuberculosis with RFLP is a very useful tool for deciphering the molecular epidemiology of tuberculosis. An international comparison of the RFLP pattern became possible with the proposal to standardize RFLP methodology using Pvu-II restricted IS-6110, and the comparison has noted some predominance of RFLP pattern in East Asia. The RFLP patterns of tuberculosis strains collected at SNUH was studied and was compared with other strains from East Asia. Method : Fifty strains of M. tuberculosis were isolated from patients who visited 'or were admitted to the SNUH in 1998. Some isolates belonging to the Beijing family were also received. After the extraction of DNA from M. tuberculosis isolates, the chromosomal DNAs were digested with Pvu-II and analyzed by the Southern blot method with DNA probe to IS6110. Result : Six strains belonged to the Beijing family. The RFLP patterns of other 9 strains were similar to each other. No statistically significant endobronchial tuberculosis, presence of underlying disease. and the province of residence were found. Conclusion : Few groups of M. tuberculosis strains from SNUH showed similar RFLP patterns, but their clinical implications are not yet clear.

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Production of Enantiopure Styrene Oxide by Recombinant Pichia pastoris carrying Double Expression cassette of Epoxide Hydrolase Gene (에폭사이드 가수분해효소 유전자의 double expression cassette 재조합 Pichia pastoris를 이용한 enantiopure styrene oxide의 제조)

  • Kim, Hee-Sook
    • Journal of Life Science
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    • v.18 no.1
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    • pp.136-142
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    • 2008
  • A recombinant Pichia pastoris carrying double expression cassette of Rhodotorula glutinis epoxide hydrolase(RgEH) gene was developed and used for preparing enantiopure (S)-styrene oxide from racemic mixture of styrene oxide. BglII restriction site of original RgEH gene (pPICZ B/RgEH #2) of previous report was mutated using PCR technique for the construction of double expression cassette containing promoter ($P_{AOX1}$), EH gene and transcription terminator ($TT_{AOX1}$) in pPICZ C vector. Double expression cassette with RgEH was inserted into the chromosomal DNA of P. pastoris. $V_{max}$ ($2.2{\mu}mol\;min^{-1}mg\;dcw^{-1}$) on (R)-styrene oxide of P. pastoris with double expression cassette was about 6-fold higher than that ($0.4{\mu}mol\;min^{-1}mg\;dcw^{-1}$) of P. pastoris with single expression cassette. For the determination of the optimal condition, the effects of detergent and temperature on the enantioselective hydrolytic activity and yield of the enantiomer were investigated. When the reaction was performed at $10^{\circ}C$ for 10 min in the presence of 0.5% Tween 20, enantiopure (S)-styrene oxide with 99.9% ee was obtained as the yield 43.4 % from 20 mM racemic sustrate.

Ten-year Clinical Study of Chorionic Villus Sampling (융모막 융모생검 511례에 대한 임상적 연구: 10년(2000-2010년)간의 경험)

  • Kim, Soo-Hyun;Shim, Sung-Han;Baek, Jong-Woo;Cha, Dong-Hyun
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.35-43
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    • 2011
  • Purpose: We evaluated indications for chorionic villus sampling (CVS), the positive predictive value of CVS for fetal chromosomal abnormalities, and the fetal loss rate after CVS at CHA Medical Center. Materials and Methods: We reviewed the medical records of 511 cases of CVS performed between 67 and 120 days of gestation for prenatal cytogenetic diagnosis from April 2000 to April 2010. Fetal karyotypes were obtained by direct and indirect culture methods. Results: The most common indications for CVS were abnormal ultrasonic findings including increased nuchal translucency (294/635, 46.3%). The positive predictive value of abnormal karyotyping according to indication for CVS was highest in cases with abnormal parental karyotypes (14/21, 66.7%). Mosaicism revealed by CVS comprised 3.1% of the sample (16/509). Amniocentesis revealed two cases of true mosaicism and 11 cases of confined placental mosaicism. The fetal loss rate within 4 weeks of the procedure was 1.2% (6/511). Conclusion: If CVS is performed by an expert clinician, it is a feasible and reliable procedure for prenatal genetic diagnosis. When CVS indicates mosaicism, the finding should be confirmed by amniocentesis to distinguish true mosaicism from confined placental mosaicism.

Rapid Detection of Enterotoxigenic Staphylococcus aureus by Polymerase Chain Reaction (중합효소 연쇄반응에 의한 식중독성 황색포도상구균의 신속한 검출)

  • Kim, Eun-Seon;Jhon, Deok-Young
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1001-1008
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    • 1996
  • Staphylococcal food poisoning is the major cause of bacterial food poisoning occurring in this country. Therefore government regulates commercial foods through Official Dictionary of Food that there should be free of enterotoxigenic Staphylococcus aureus in Korean rice cakes, bread, and a box lunch. Since at least 5 days are required to identify the S. aureus by the official method in the Dictionary it is difficult to prevent the food poisoning and the investigation of the outbreaks. In this report an improved determination method of the S. aureus has been developed using polymerase chain reaction (PCR) technique. Sense and antisense primers for specific amplification of genes encoding staphylococcal enterotoxins were designed and synthesized for the PCR. Rapid chromosomal DNA isolation method was also developed from S. aureus using lysostaphin. The PCR condition was developed as follows. Reaction solution $(50\;{\mu}l)$ consisted of target DNA $2\;{\mu}l$ (about 20ng), 10X buffer $5\;{\mu}l$, primer 100pmole, dNTP (10 mM) $4\;{\mu}l$ and Taq DNA polymerase 2.5 unit in a thin-wall tube. Operation condition of the PCR was 5 min pre-denaturation at $94^{\circ}C$, 15 sec denaturation at $94^{\circ}C$, 15 sec annealing at $50^{\circ}C$, 20 sec extension at $72^{\circ}C$, and 5 min post-extension at $72^{\circ}C$, and 30 cycles of denaturation-annealing- extension. Using the PCR with Perkin Elmer GeneAmp PCR system 2400, types of enterotoxigenic S. aureus could be identified from Ddok or bread in a day.

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