• 제목/요약/키워드: DNA and protein patterns

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Intratesticular Injection of Hypertonic Saline: Non-Invasive Alternative Method for Animal Castration Model

  • Kwak, Byung Kuk;Lee, Sung-Ho
    • 한국발생생물학회지:발생과생식
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    • 제17권4호
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    • pp.435-440
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    • 2013
  • Previous studies, including our own, have demonstrated that the intratesticular injection of hypertonic saline (20%) decreased serum testosterone level which was similar to the surgical castration in the rat, showing the state of chemical castration. In the present study, we further verify the efficacy of this less invasive method as an alternative of surgical orchidectomy in the andrological field. Sterilized 20% saline was directly injected into the adult male rats (750 ${\mu}l$ per testis). The tested rats were divided into 3 groups including intact group (intact), orchidectomy group (ORX) and saline injection group (SAL) after bilateral orchidectomy was performed at the same day of injection. All rats were sacrificed at 4 weeks after injection. The reproductive organs (testes, epididymis, seminal vesicles and prostates) were collected and used for DNA and protein pattern analyses. Also, patho-histological studies on the testes were performed. In contrast to the intact group, similar DNA damages of testis and seminal vesicle were appeared in ORX group and SAL group. The DNA degradations seemed to be the results of necrosis rather than apoptosis. In the protein pattern analysis, all the testing tissues exerted similar patterns in the ORX group and the SAL group compared to the those of intact group. Patho-histological studies revealed that severe degenerative changes in testicular seminiferous tubules and massive infiltration of immune cells in SAL group. The present study confirmed that direct injection of hypertonic saline into the testis caused the equivalent biochemical changes in the accessory sex organs as shown in the orchidectomized animals. These results suggest that hypertonic saline injection model could be a useful castration model which can substitute for surgical castration when its safety is secured through further study in the future.

감귤에서 분리한 Metallothionein 유전자의 발현분석 및 게놈 DNA (Expression Patterns and Isolation of Genomic DNA of a Metallothionein-like Gene from Citrus (Citrus unshiu Marc. cv. Miyagawa))

  • 김인중
    • 식물조직배양학회지
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    • 제28권5호
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    • pp.231-237
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    • 2001
  • Differential screening을 통해 Moriguchi등 (1998)이 분리한 유전자와 상동성을 나타내는 CitMT45 유전자의 cDNA를 분리하였다. 본 실험에서 분리한 cDNA는 Moriguchi등 (1998)이 분리한 cDNA에 비해 긴 3' UTR을 가지고 있었다. 잎과 과피, 과육에서 CitMT45 유전자의 발현분석을 northern blot을 통해 수행한 결과, 발달단계에 따라 증가하는 비슷한 앙상을 관찰할 수 있었으나, 과육, 과피, 잎의 순으로 그 발현 양이 많았다. 이들의 발현조절에 대한 정보를 얻기 위해 게놈 DNA를 분리한 결과, CitMT45 게놈 구조는 3개의 exon과 2개의 intron으로 구성되어 있었고, primer extension 분석을 통해 CitMT45 유전자의 발현은 3개의 부위에서 개시되고 있음을 알 수 있었다. 전사개시부위의 5'upstream 지역에서 TATA box와 CCAAT box뿐만 아니라, 금속이온과 온도변화에 의한 조절에 중요한 부위로 알려진 cis-element를 발견하였다.

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연쇄 구균의 세포벽 단백질이 L929 세포의 DNA합성에 미치는 영향 및 SDS-PAGE 양상에 관한 연구 (THE EFFECTS OF CELL WALL PROTEINS OF STREPTOCOCCUS SPP. ON DNA SYNTHESIS OF L929 CELLS AND THEIR SDS-PAGE PATTERNS)

  • 이세종;임미경
    • Restorative Dentistry and Endodontics
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    • 제20권1호
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    • pp.71-95
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    • 1995
  • Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

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인공 산성비가 중요 농작물의 생육에 미치는 영향 (The Effect of Simulated Acid Rain on the Growth of Important Crops)

  • 허흥욱;허만규
    • 한국환경과학회지
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    • 제7권2호
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    • pp.123-132
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    • 1998
  • The experiment was performed to elucidate the effect of simulated acrid rain on growth of the seven crops(caucus carota vu sativa DC. , Fagopyrum esculenkm Moench, Brassica cmpespis subsp. napus var. pekinensis Makino , Raphuus satlws vu. hortensis for. acnMormis Makino, Brassica dbogjabra Bailey, Caphslcum mum L., and Perilla frutescens Britton). The pH levels of simulated acid rain ranged pH 3.1, 3.6, 4.1, 4.6, 5.1 and 5.6. The germination 10 each crop was influenced from stimulated acrid rain except buckwheat and kale. A general decrease of growth in crops was observed with Increasing pH concentrations. The pattern of soluble protein was observed a tendency to decrease from acidic pH eradlents. According to acidity, total DNA contents of each crop was showed a definite reduction. In conclusion, plant growth was stimulated decreasln and the chanties of total Protein Patterns and BNA contents extracted from leaves trended with stimulated acrid rain was showed seriously.

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Hc nuclear polyhedrosis virus DNA 제한효소절편의 molecular cloning 과 외래 유전자 발현 (Molecular cloning and foreign gene expression of restriction endonuclease fragments of the Hc nuclear polyhedrosis virus DNA)

  • 이근광
    • 한국어병학회지
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    • 제8권1호
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    • pp.31-36
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    • 1995
  • HcNPV DNA genome 을 제한효소 EcoRI 으로 절단하여 그들의 일부 절편을 pUC8 vector 에 cloning 한 후 E. coli JM 83 세포에 형질 전환시켰다. 이 결과 24 개의 EcoRI 절편중 12 개의 절편이 cloning 되었다. 이들 제조합체중 4 개는 eNP-O, eNP-Q, eNP-R, eNP-S 라 명명하였다. 또한 이들 제조합체의 외래 유전자 발현을 SDS-PAGE 에 의해 단백질 패턴을 분석하였다. 그 결과 제조합체 eNP-O, eNP-Q, eNP-R 에서는 E. coli JM 83 숙주세포의 단백질 밴드와 비교하여 다른 분자량을 갖는 밴드가 나타났다.

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Bombyx mori세포주와 Spodoptera frugiperda세포주의 분자생물학적 표식자 (The Molecular Biological Marker in Bombyx mori and Spodoptera frugiperda Cells)

  • 진병래;제연호;강석권
    • 한국잠사곤충학회지
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    • 제38권1호
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    • pp.53-56
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    • 1996
  • 곤충세포주로 널리 이용되고 있는 Sf 세포주와 Bm 세포주의 분자생물학적 표식자를 탐색하기 위하여, 총 세포 단백질의 SDS-PAGE와 genomic DNA를 RAPD(Random Amplification of Polymorphic DNA) 방법으로 분석하였다. 그 결과, 총 세포 단백질 및 genomic DNA, 패턴에서 두 세포주를 뚜렷하게 구별할 수 있는 밴드를 탐색하였으며, 이들은 유용한 분자 생물학적 표식자로 이용될 수 있을 것이다.

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새로운 곤충병원성 Bacillus thuringiensis subsp. aizawai 균주의 유전학적 특성 (Genetic characteristics of the novel insect pathogenic Bacillus thuringiensis subsp. aizawai strain)

  • 서미자;윤영남;유용만;김기수
    • 농업과학연구
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    • 제41권4호
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    • pp.351-359
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    • 2014
  • For identifying the plasmid DNA coding cry gene of Bacillus thuringiensis subsp. aizawai KB098 with high insecticidal activity against Spodoptera exigua, mutant isolates with no crystal protein were produced by $42^{\circ}C$ incubation condition and then mutant plasmid DNA band patterns were compared with those of KB098. KB098 isolates had 4 cry genes, cry1Aa, cry1Ab, cry1C, cry1D, and also had been found seven plasmid DNA. Though the SDS-PAGE experiment, it was confirmed that mutant didn't produce 130~145kDa protein band involved in bipyramidal shape crystal. Also, five mutant isolates had no cry genes coding plasmid DNA in PCR. In result of comparison the plasmid DNA of KB098 and 5 mutant isolates, only 1 plasmid DNA band was left out in mutant plasmid DNA pattern, so that the missing band was extracted from the gel. The missing(disappeared) plasmid DNA was the largest molecular size among the 7 plasmid DNA of KB098 and it was also confirmed this plasmid DNA had all 4 cry genes through PCR.

Isolation and Characterization of Chlorella Virus from Fresh Water in Korea and Application in Chlorella Transformation System

  • Park, Hye-Jin;Yoon, Hong-Mook;Jung, Heoy-Kyung;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • 제21권1호
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    • pp.13-20
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    • 2005
  • Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.

Classification of Colon Cancer Patients Based on the Methylation Patterns of Promoters

  • Choi, Wonyoung;Lee, Jungwoo;Lee, Jin-Young;Lee, Sun-Min;Kim, Da-Won;Kim, Young-Joon
    • Genomics & Informatics
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    • 제14권2호
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    • pp.46-52
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    • 2016
  • Diverse somatic mutations have been reported to serve as cancer drivers. Recently, it has also been reported that epigenetic regulation is closely related to cancer development. However, the effect of epigenetic changes on cancer is still elusive. In this study, we analyzed DNA methylation data on colon cancer taken from The Caner Genome Atlas. We found that several promoters were significantly hypermethylated in colon cancer patients. Through clustering analysis of differentially methylated DNA regions, we were able to define subgroups of patients and observed clinical features associated with each subgroup. In addition, we analyzed the functional ontology of aberrantly methylated genes and identified the G-protein-coupled receptor signaling pathway as one of the major pathways affected epigenetically. In conclusion, our analysis shows the possibility of characterizing the clinical features of colon cancer subgroups based on DNA methylation patterns and provides lists of important genes and pathways possibly involved in colon cancer development.

Expression of major piroplasm protein(p33)of Theileria sergenti (Korean isolate) and its immunogenicity in guinea pigs

  • Kang, Seung-Won;Kweon, Chang-Hee;Choi, Eun-Jin;Yoon, Yong-Dhuk
    • Parasites, Hosts and Diseases
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    • 제37권4호
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    • pp.277-283
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    • 1999
  • To investigate the development of a subunit vaccine against theileriosis in cattle, the DNA fragments encoding piroplasm surface protein (p33) of Theileria sergenti of a Korean isolate were expressed in baculoviruses. The expressed p33 was characterized by indirect fluorescent antibody (IFA) and western blotting analysis. The expression of p33 was mainly detected on the surface of infected Sf21 cells by IFA. The immunoblotting analysis revealed the presence of a same molecular weight protein band of p33. The antigenicity of expressed polypeptide was further examined through the inoculation of a guinea pig. The sera of guinea pigs immunized with p33 expressed cell Iysate showed similar fluorescent antibody patterns and reacted with the same molecular weight protein of T. sergenti in immunoblotting analysis, thus indicating that this protein can be a promising candidate for a subunit vaccine in the future.

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