• 제목/요약/키워드: Cell Transformation

검색결과 622건 처리시간 0.035초

Improvement of Transformation Efficiency Through In Vitro Methylation and SacII Site Mutation of Plasmid Vector in Bifidobacterium longum MG1

  • Kim, Jin-Yong;Wang, Yan;Park, Myeong-Soo;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권6호
    • /
    • pp.1022-1026
    • /
    • 2010
  • The different cleavage patterns of pYBamy59 plasmid isolated from E. coli $DH5{\alpha}$ and B. longum MG1 by the cell extract of B. longum MG1 suggested that the main reason for its low transformation efficiency was related to the restriction modification (R-M) system. To confirm the correlation between the R-M system and transformation efficiency, in vitro methylation and site-directed mutagenesis were performed in pYBamy59. Sequence analysis of pYBamy59 fragments digested by the cell extract of B. longum MG1 revealed that all fragments were generated by restriction of the sequence recognized by SacII endonuclease. When pYBamy59 from E. coli was methylated in vitro by CpG or GpC methyltransferase, it was protected from SacII digestion. Site-directed mutagenesis, which removed SacII sites from pYBamy59, or in vitro methylation of pYBamy59 showed 8- to 15-fold increases in the transformation efficiency over intact pYBamy59. Modification of the SacII-related R-M system in B. longum MG1 and in vitro methylation in pYBamy 59 can improve the transformation efficiency in this strain. The results showed that the R-M system is a factor to limit introduction of exogenous DNA, and in vitro modification is a convenient method to overcome the barrier of the R-M system for transformation.

C-TRANSFORMATIONS ON OPEN 3-CELLS

  • Lee, Joo Sung
    • Korean Journal of Mathematics
    • /
    • 제9권2호
    • /
    • pp.75-82
    • /
    • 2001
  • In this paper, we show the existence of open subsets U of $S^3$ which admit a C-transformation onto the interior of $B^3$. Let U be an open set which is homeomorphic to the interior of $B^3$. Then, we prove that if U has a pseudo general polyhedral prime end structure then U admits a C-transformation.

  • PDF

사람 유래의 MCF10A, Chang liver및 HaCaT 세포의 소핵형성 및 세포형질전환에 미치는 2,3,7,8-Tetrachlorodibenzo-p-dioxin의 영향

  • 엄미옥;박미영;김종원;박미선;한의식;오혜영;정해관
    • 한국환경성돌연변이발암원학회지
    • /
    • 제24권2호
    • /
    • pp.91-98
    • /
    • 2004
  • Although 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) is a powerful carcinogen in several species, limited model system exist to study carcinogenicity of this compound at cellular level. To enhance our under-standing of carcinogenicity of TCDD at cellular level, we investigated micronucleus (MN) frequency as a index of genetic toxicity and whether TCDD can transform the human cells in culture. Normal human cell lines, skin keratinocyte HaCaT, Chang liver and breast MCF10A cells were used. TCDD did not affect the cell viability of the Chang liver, HaCaT and MCF10A cells. The frequency of micronucleus was increased after treatment of TCDD for 24hr in Chang liver and HaCaT cells, but not changed in MCF10A cells. And we observed putative transformed cells in Chang liver cells exposed to 1 $\mu$M TCDD for 2 weeks. The putative transformed cells were also observed in HaCaT cells with subsequent exposure to TCDD (0.1, 1, 10, 100 nM) for 2 weeks after initial exposure to MNNG, but not observed in MCF10A cells. Collectively, these results indicate that the ability of TCDD to induce micronuclei may be involved in cellular transformation of Chang liver and HaCaT cells. Our putative TCDD-transformed cells of Chang liver and HaCaT are expected to provide a clue to the elucidation of TCDD-induced transformation pathway.

  • PDF

방사선 조사 없이 속발한 거대 세포종의 악성 전환 (Malignant Transformation of Giant Cell Tumor Not Associated with Radiotherapy)

  • 이상훈;오주한;유광현;서성욱;안준환;김한수;임수택
    • 대한골관절종양학회지
    • /
    • 제8권1호
    • /
    • pp.12-19
    • /
    • 2002
  • 목적 : 거대 세포종이 방사선 조사 없이 악성 전환하는 것은 매우 드문 경우로 저자들이 경험한 4례를 분석하여, 흔하지 않은 악성 전환된 거대 세포종의 진단과 치료의 방침을 제시하는데 도움이 되고자 한다. 대상 및 방법 : 1985년 9월부터 2001년 1월까지 본원 정형외과에 내원하여, 수술적 치료를 시행했던 거대 세포종 112례 중 방사선 조사 없이 악성화한 4례를 대상으로 하였으며, 악성전환 후 평균 추시 기간은 2.4년(최저 1.3년, 최고 4년)이었다. 결과 : 거대 세포종의 진단 후 악성 전환 때까지의 기간은 평균 6.9년(최저 2.2년, 최고 13.5년)이었고, 근위 상완부 연부 조직, 근위 대퇴골, 원위 대퇴골, 근위 경골에 각각 1례씩 발생하였다. 3례에서는 골 육종으로, 1례에서는 악성 섬유성 조직구종으로 악성 전환이 일어났으며, 국소 재발은 1례, 폐 전이는 골 육종으로 악성 전환한 3례에서 모두 관찰되었다. 결론 : 방사선 조사 없이 속발한 거대 세포종의 악성 전환을 진단하기 위해서는 병리 조직의 철저한 채취가 매우 중요한 요소라 사료되며, 방사선 소견상 악성 전환이 의심될 때는 부적절한 절제술을 시행하기보다는 절개 생검을 먼저 시행한 후 치료 방침을 정해야 할 것으로 사료된다.

  • PDF

Immunohistochemical study on the expression of matrix metalloproteinase 2 and high-risk human papilloma virus in the malignant progression of papillomas

  • Lee, Ho-Jin;Kim, Jin-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • 제39권5호
    • /
    • pp.224-230
    • /
    • 2013
  • Objectives: Papilloma frequently develops as a benign tumor of the head and neck area, but its potential for malignant transformation has yet to be studied. This study aims to provide basic information for papillomas using the immunohistochemical staining of matrix metalloproteinase 2 (MMP-2) and human papilloma virus (HPV) 16 and 18. Materials and Methods: To evaluate the malignant transformation of papillomas, the selected tissue samples were serially diagnosed with pre-cancerous papilloma (with epithelial dysplasia, pseudo-epitheliomatous hyperplasia) or malignant lesion (squamous cell carcinoma, SCC) after the first diagnosis (squamous papilloma, inverted papilloma). The selected tissues were stained with an antibody to MMP-2 and HPV 16-E7, HPV 18-L1. A statistical analysis was performed according to each transformation step. Results: The epithelial layer of papilloma and pre-cancerous papilloma lesions had a similar MMP-2 expression, but that of the malignant lesion had a significantly increased MMP-2 expression. HPV 16 and 18 infection rates were 28.6%, 33.3% and 63.6% in papillomas, pre-cancerous papilloma lesions, and SCC. Conclusions: A relatively high MMP-2 expression and HPV 16 or 18 infection of papillomas may be associated with early events in the multistep processes of malignant transformation of papillomas.

Solvent effect on enzymatic steroid transformation

  • Kim, Doo-Ha;Lee, S.B.;Ryu, D.Y.
    • Archives of Pharmacal Research
    • /
    • 제3권1호
    • /
    • pp.1-6
    • /
    • 1980
  • As part of our endeavor to increase the productivity of steroid by enzymatic transformation of corticosteroids, attempts have been made to increase the solubility of steroids by using some organic solvents. When the solubility of steroids is the rate limiting factor in the steroid transformation, it was found that the use of solvents significantly improved the yield. Hydrocorisone as a substrate and 3-ketosteroid .DELTA.$^{1}$ dehydrogense as an immobilized whole cell enzyme were employed as the model system for this study. It was found that the yield of product, prodnisolone, goes through a maximum with an increase in the solvent concentration. At a high solvent concentration, the solvent showed a toxic effect and it causes a decrease in the product yield by the second order inhibition mechanism. Among the solvents evaluated, methanol and ethanol were found to be the best. These alcohols are not only good solvents but also showed minimal toxic effect. Based on the experimental results, it was concluded that the productivity of steroid can be increased by usign well selcted solvents systems for the enzymatic transformation of steroids.

  • PDF

Effect of Nisin against Clostridium botulinum During Spore-to-Cell Transformation

  • Chung, Yoon-Kyung;Yousef, Ahmed E.
    • Preventive Nutrition and Food Science
    • /
    • 제12권4호
    • /
    • pp.259-266
    • /
    • 2007
  • It has been proposed that the mode of action of nisin against vegetative cells and spores of Clostridium botulinum is different. However, clear explanation is not available. Therefore, nisin action against vegetative cells and spores of C. botulinum was investigated in this study. Nisin was added at various stages of spore-to-vegetative cell transition and changes to sensitivity to the bacteriocin were observed. Different nisin preparation (Nisaplin or pure nisin) was compared for their activity against different stages of spore transformation of C. botulinum ATCC 25763. Germination was measured by determining loss of heat resistance and observing phase darkening of spores under phase-contrast microscope. Nisin acted bactericidally against vegetative cells, but acted sporostatically against spores of C. botulinum under the same concentration. This bactericidal and sporostatic action of nisin was dependent on the concentration of nisin used. Presence of nisin during spore activation by heat increased subsequent phase darkening and germination rates. However, nisin inhibited the germination and the outgrowth, when it was added after heat activation stage. Findings from this study suggest that the time of addition of nisin is very important for the effective control of spores during the heating process of foods. In addition, it may be possible to apply nisin at the stage of processing that coincides with the most sensitive stage of spore transformation.

Transformation of a Filamentous Fungus Cryphonectria parasitica Using Agrobacterium tumefaciens

  • Park, Seung-Moon;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제9권3호
    • /
    • pp.217-222
    • /
    • 2004
  • As Agrobacterium tumefaciens, which has long been used to transform plants, is known to transfer T-DNA to budding yeast, Saccharomyces cerevisiae, a variety of fungi were subjected to the A. tumefaciens-mediated transformation to improve their transformation frequency and feasibility. The A. tumefaciens-mediated transformation of chestnut blight fungus, Cryphonectria parasitica, is performed in this study as the first example of transformation of a hardwood fungal pathogen. The transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator, as a selectable marker, led to the selection of more than 1,000 stable, hygromycin B-resistant transformants per 1${\times}$10$\^$6/ conidia of C. parasitica. The putative transformants appeared to be mitotically stable. The transformation efficiency appears to depend on the bacterial strain, age of the bacteria cell culture and ratio of fungal spores to bacterial cells. PCR and Southern blot analysis indicated that the marker gene was inserted at different chromosomal sites. Moreover, three transformants out of ten showed more than two hybridizing bands, suggesting more than two copies of the inserted marker gene are not uncommon.

Identification of Plant Factors Involving in Agrobacterium-mediated Plant Transformation

  • Nam, Jaesung
    • 식물조직배양학회지
    • /
    • 제27권5호
    • /
    • pp.387-393
    • /
    • 2000
  • The process by which Agrobacterium tumefaciens genetically transforms plants involves a complex series of reactions communicated between the pathogen and the plants. To identify plant factors involved in agrobacterium-mediated plant transformation, a large number of T-DNA inserted Arabidopsis thaliana mutant lines were investigated for susceptibility to Agrobacterium infection by using an in vitro root inoculation assay. Based on the phenotype of tumorigenesis, twelve T-DNA inserted Arabidopsis mutants(rat) that were resistant to Agrobacterium transformation were found. Three mutants, rat1, rat3, and rat4 were characterized in detail. They showed low transient GUS activity and very low stable transformation efficiency compared to the wild-type plant. The resistance phenotype of rat1 and rats resulted from decreased attachment of Agrobacterium tumefaciens to inoculated root explants. They may be deficient in plant actors that are necessary for bacterial attachment to plant cells. The disrupted genes in rat1, rat3, and rat4 mutants were coding a arabinogalactan protein, a likely cell wall protein and a cellulose synthase-like protein, respectively.

  • PDF