• 제목/요약/키워드: SV40

검색결과 216건 처리시간 0.022초

SV 40 바이러스가 유도한 DNA 합성효소의 특성에 대한 연구 (Characterizations of DNA-polymerases Induced by SV40 Virus Infection of African Green Monkey Kidney Cells (AGMK))

  • 강현삼
    • 미생물학회지
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    • 제14권3호
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    • pp.135-145
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    • 1976
  • Confluent AGMK cells were infected by large plaque SV40 virus. Levels of DNA polymeras $({\alpha}\;and\;{\beta})$ were measured in the cytoplasm and the cell nucleus. The activities of DNA $polymerase-{\alpha}$ which found in both the cell nucleus and the cytoplasm were increased approximately eight folds at 48 hours after infection of SV40 virus. Only insignificant but constant amounts of DNA $polymerase-{\beta}$ were found either in the nucleus of the SV40 infected cell or of the uninfected cell. The characteristics of the SV40 virus induced DNA polymerases were compared with that of the uninfected cellular DNA polymerase in regard of the effects of pH, salt concentration, NEM concentration and temperature on those enzyme activities. No differential effect was found between both enzymes. Endouclease activities wre examined in the purified DNA $polymerase-{\alpha}\;and\;{\beta}$. The low level of endonuclease activity which might cut SV40 DNA 1 at one site was observed in the DNA $polymerase-{\alpha}$ whereas high but nonspecific endonuclease activities were found in the DNA $polymerase-{\beta}$.

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SV 40 DNA를 이용한 포유동물의 유전자 운반체 개발 (Construction of an expression vector with SV40 DNA in a mammalian cell)

  • 정민혜;김상해;전희숙;노현모
    • 미생물학회지
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    • 제25권3호
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    • pp.165-172
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    • 1987
  • An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1, 118bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVML-TKp. To test the expression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kinase gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK. These plasmids, pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.

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Impact of SV40 T antigen on two multiple fission microalgae species Scenedesmus quadricauda and Chlorella vulgaris

  • Gomaa, Ahmed E.;Yang, Seung Hwan
    • International journal of advanced smart convergence
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    • 제7권1호
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    • pp.48-63
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    • 2018
  • The combination of Simian Virus40 (SV40)'s large T antigen with its replication origin is commonly used in molecular studies to enhance the expression of heterogeneous genes through multiplying the plasmid copy number. There are no reports related to the impact of the SV40 T antigen on plant, multiple fissional, cell-type. This study explores the response of two multiple-fission microalgal cells, Scenedesmus quadricauda and Chlorella vulgaris, to the expression of the T-antigen, with aim of applying SV40 T-antigen to increase the expression efficiency of foreign genes in the two species. Different levels of low-expression have been constructed to control the expression of SV40 T antigen using three heterogenous promoters (NOS, CaMV35S, and CMV). Chlorella cultures showed slowdown in the growth rate for samples harboring the T antigen under the control of CaMV35S and CMV promoters, unlike Scenedesmus cultures which showed no significant difference between samples and could have silenced the expression.

원숭이 신장 세포에서 M13 DNA에 의한 SV40 DNA 복제 억제 현상에 대하여 (Inhibition of SV40 DNA replication in simian cell by bacteriophage M13 DNA sequences)

  • 김연수;구용의;강현삼
    • 미생물학회지
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    • 제26권3호
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    • pp.162-166
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    • 1988
  • Bacteriophage M13 DNAs carrying the wild type or base substituted SV40 DNA replication origins were used for replication assay. In vivo and in vitro assay with African green monkey cell line COS-1 showed that the replication of M13-SV40 recombinant DNAs was restricted like a pBR322 SV40 recombinant DNA(Lusky and Botchan, 1981). Furthermore, recombinant phage DNAs isolated from the transfected siminan cells subsequently show a reduced ability to retransform E. coli. But pATSV-W(Kim et al., 1988) was replicated in COS-1 cells normally. We think that a poison sequence may exist on bacteriophage M13 DNA like pBR322.

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Establishment and characterization of an immortalized human dermal papilla cell line

  • Shin, Seung-Hyun;Park, Sang-Yoon;Kim, Moon-Kyu;Kim, Jung-Chul;Sung, Young-Kwan
    • BMB Reports
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    • 제44권8호
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    • pp.512-516
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    • 2011
  • Establishment of immortalized human dermal papilla cells (DPCs) retaining the characteristics of DPCs would be a great help for hair researchers. We recently established a simian virus 40T (SV40T)-transformed human DP cell line (SV40TDPC). However, the cell line senesced around passage 25 and ceased proliferation. In this study, we introduced the human telomerase reverse transcriptase (hTERT) gene into SV40T-DPC and established an immortalized human DP cell line. The cell line, SV40T-hTERT-DPC, did not induce tumors when inoculated into nude mice. SV40T-hTERT-DPC maintained morphology of early passage DPCs, expressed markers of DPCs, and retained responses to Wnt/${\beta}$-catenin and bone morphogenic protein (BMP) signaling pathways known to be required for hair-inducing activity of DPCs. The data strongly suggest that SV40T-hTERT-DPC retains many characteristics of human DPCs in vivo without malignant transformation.

소성장호르몬 유전자의 조직 특이성 발현에 미치는 바이러스 engancer의 영향 (Effect of Viral Enhancers on the Tissue-Specific Expression of Bovine Growth Hormone Gene)

  • 박계윤;김수미;노정혜
    • 미생물학회지
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    • 제27권2호
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    • pp.85-91
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    • 1989
  • 조직특이적 및 일반적 유전자 발현에 미치는 SV40와 murine cytomegalovirus (MCMV) enhancer의 영향을 조사하였다. 이를 위하여 chloramphenicol acetyl transferase (CAT) 유전자의 아래쪽에 이들 enhancer들을 삽입한 재조합 플라스미드 들을 제조하였다. 원숭이세포(CV1PO)와 HeLa 세포에 이들 플라스미드들을 이입시킨 후, CAT 유전자가 발현되는 정도를 조사하였다. Enhancer가 없는 플라스미드에 비해 SV40와 MCMV enhancer는 CAT의 발현을 각각 20배와 150배로 가중시켰다. CAT 유전자의 앞에 있는 SV40 프로모터를 2.2kbp의 소성장호르몬(bGH) 유전자의 조절부위로 치환한 경우는 enhancer가 있어도 전혀 CAT 의 말현이 검출되지 않았다. 조절부위을 230 bp 로 짧게 하여 치환한 경우는, SV 40 enhancer 가 있을 때, CAT의 말현이 매우 증가하였다. 이와는 내조적으로 더 강한 MCMV enhancer는 bGH 특이적인 발현을 별로 증가시키지 못하였다.

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새로운 항암제 DA-125의 유전자 복제 억제 기작 (Inhibitory Mechanism of a New Antitumor Agent DA125 on DNA Replication)

  • 이상광;김도진;오유택;이상득;우은란;신차균
    • 약학회지
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    • 제43권5호
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    • pp.623-628
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    • 1999
  • DA-125, a new antitumor agent, was compared with adriamycin, a known DNA intercalator, in terms of inhibitory mechanism of DNA replication by using replicating simian virus 40 (SV40) genome in vivo. In analyzing the SV40 DNA replication intermediates present in cells treated with DA-125, it was not observed to accumulate B-dimers of SV40 DNA which are prominent in adriamycin-treated cells. However, treatment with DA-125 induced dose-dependent formation of DNA-topoisomerase complex which is characteristic of topoisomerase poisons. In addition, DA-125 showed more efficient in inhibiting SV40 DNA replication than adriamycin. Therefore, on the basis of this observation, we suggest that DA-125, a derivative of adriamycin, inhibits DNA replication by blocking topoisomerase activity as a toposomerase poison although adriamycin blocks topoisomerase activity as a DNA intercalator.

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X선 촬영에서 조사야 크기에 따른 산란선량의 변화 (Change of the Scattered Dose by Field Size in X-ray Radiography)

  • 최성관
    • 한국콘텐츠학회논문지
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    • 제13권3호
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    • pp.198-203
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    • 2013
  • 본 연구에서는 손, 머리, 복부 등에 대한 X선 촬영 시행 시 조사야 크기를 최적화할 경우와 최대화할 경우 검사목적부위로부터 30cm 거리에서의 X선 산란선량이 각각 어느 정도인지를 알아보았다. 그 결과 손, 머리, 복부 등에 대한 X선 산란선량은 첫째, 소인촬영의 경우 조사야 크기를 최적화하였을 때 각각 $0.08{\mu}Sv$, $4.39{\mu}Sv$, $5.56{\mu}Sv$로 나타났고, 조사야 크기를 최대화하였을 때 각각 $0.58{\mu}Sv$, $33.47{\mu}Sv$, $35.93{\mu}Sv$로 나타났으며, 둘째, 성인촬영의 경우 조사야 크기를 최적화하였을 때 각각 $0.40{\mu}Sv$, $14.51{\mu}Sv$, $18.86{\mu}Sv$로 나타났고, 조사야 크기를 최대화하였을 때 각각 $2.78{\mu}Sv$, $107.40{\mu}Sv$, $117.52{\mu}Sv$로 나타났다(P<0.001). 결론적으로, X선 촬영 시 조사야 크기를 필요한 만큼만으로 최대한 줄여주어 최적화시켰을 때에 최대화 시켰을 때보다 피사체 주변의 X선 산란선 발생량은 약 6~7배 정도 감소하였다.

SV40 바이러스로 형질전환된 사람종양세포의 특성 (Proporties of SV4O-transformed Human Cells)

  • 최경희;홍승환
    • 한국동물학회지
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    • 제31권1호
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    • pp.49-55
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    • 1988
  • SV80와 같은 SV40로 형질전환된 사람세포는 종양을 일으킬 수 있는 능력을 가지고 있으나 면역기구인 흉선이 없는 누드마우스에서는 거부반응을 일으켜 종양을 일으키지 않는다. 그러나, 예외적으로 WI18/VA-2세포는 누드마우스에서 종양을 일으키며 이에서 얻는 두클론중 NW18C11은 종양을 일으키나 NW18C12는 종양을 일으키지 않는다. 본 실험에서는 이들 두 클론의 차이점들을 조사하였다. 실험결과, NW18C11은 NW18C12보다 더 많은수의 SV40 sequence를 포함하고 있음을 southern blot방법을 통해 확인하였으며 또한 immunofluoresce와 immunoprecipitation방법을 사용하여 두 클론 모두 정상크기의 SV40유전자산물인 large T와 small t 단백질을 생성함을 확인하였다. 한편 두 클론내에 포함되어 있는 바이러스유전자가 비형질전환새포로 하여금 생체내에서 악성종양 형성능력을 획득하도록 형질전환시킬수 있는지 확인하기 위해 두 클론의 DNA를 추출하여 마우스 NIH3T3세포에 주입시켜 형질전환된 세포를 선별하였다. 이 세포들은 모두 large T단백질을 생성하였으며 누드마우스에서 종양을 일으켰다. 이들 결과로써 NW18C12세포의 형질전환능은 완전하며, 이 세포가 누드마우스에서 거부반응에 기인하는 것으로 생각된다.

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SV 40 Promoter를 갖는 Plasmid에 의한 NIH3T3 섬유아세포의 형질전환 (Transforming Capacity of the Plasmid Containing SV40 Promoter in NIH3T3 Fibroblast Cells)

  • 이영환;김광식;서용택;김용웅;박남용;황태주
    • 미생물학회지
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    • 제27권1호
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    • pp.10-15
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    • 1989
  • Mammalian cell 연구에 쓰기 위해 개발된 SV 40 transcriptional promoter를 함유하는 pKOneo plasmid를 발암 유전인자 연구에 쓰이는 NIH3T3 쥐 세포에 stable transfection 시켜 7개의 sub clones 얻었으며, 이 subclones이 갖는 세포 형질전환에 관한 여러가지 성질을 조사하였다. 실험결과에 따르면 stable transfection 후 세포 염색체에 삽입된 pKOneo plasmid 자체만으로도 NIH3T3 세포의 형질전환을 크게 일으키는 것으로 사료되었다.

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