• Title/Summary/Keyword: K-L transformation

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Cells Transformed by PLC-Gamma 1 Overexpression are Highly Sensitive to Clostridium difficile Toxin A-Induced Apoptosis and Mitotic Inhibition

  • Nam, Hyo-Jung;Kang, Jin-Ku;Chang, Jong-Soo;Lee, Min-Soo;Nam, Seung-Taek;Jung, Hyun-Woo;Kim, Sung-Kuk;Ha, Eun-Mi;Seok, Heon;Son, Seung-Woo;Park, Young-Joo;Kim, Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.50-57
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    • 2012
  • Phospholipase C-${\gamma}l$ (PLC-${\gamma}l$) expression is associated with cellular transformation. Notably, PLC-${\gamma}$ is up-regulated in colorectal cancer tissue and breast carcinoma. Because exotoxins released by Clostridium botulinum have been shown to induce apoptosis and promote growth arrest in various cancer cell lines, we examined here the potential of Clostridium difficile toxin A to selectively induce apoptosis in cells transformed by PLC-${\gamma}l$ overexpression. We found that PLC-${\gamma}l$-transformed cells, but not vector-transformed (control) cells, were highly sensitive to C. difficile toxin A-induced apoptosis and mitotic inhibition. Moreover, expression of the proapoptotic Bcl2 family member, Bim, and activation of caspase-3 were significantly up-regulated by toxin A in PLC-${\gamma}l$-transformed cells. Toxin A-induced cell rounding and paxillin dephosphorylation were also significantly higher in PLC-${\gamma}l$-transformed cells than in control cells. These findings suggest that C. difficile toxin A may have potential as an anticancer agent against colorectal cancers and breast carcinomas in which PLC-${\gamma}l$ is highly up-regulated.

Selection of Suitable Varieties of Carnation (Dianthus caryophyllus L.) and Optimization of Culture Conditions for Efficient Tissue Culture (효율적 조직배양체계 확립을 위한 카네이션 품종 선발 및 배양조건 설정)

  • Kang, Chan-Ho;Han, Bum-So;Han, So-Gon;Kown, Sung-Hwan;Song, Young-Ju
    • Korean Journal of Plant Resources
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    • v.24 no.2
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    • pp.121-129
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    • 2011
  • As the molecular breeding was progressed, many plant transformation techniques were attained for improving transformation accuracy and used to produce useful transgenic plants. Day by day, new varieties were developed so new transformation techniques required for these newly developed varieties. Carnation (Dianthus caryophyllus L.) is a popular and economically important ornamental plant, all over the world. Keeping this in view, we selected 18 varieties of D. caryophyllus L. commonly available in the market and did optimization of culture conditions for more efficient tissue culture and to get higher number of plants via micro-propagation. Four varieties namely Yellowdotcom, Jakarta, Belmonte, Polartessino etc. were selected for organ culture studies from single cell line. The optimum growth was recorded in the MS media supplied with sucrose 3%, NAA 1.0 mg/L and TDZ 1.0 mg/L. except Belmonte, in which, BA 1.0 mg/L was found to be the best combination, in place of TDZ, rest ingredients were same. The most efficient coagulating agent used to obtain higher number of plant from callus was phytagel 0.3%. The most effective explant for higher shoot formation was stem in which 80.2% shoot formation was recorded. It also reduced culture periods by 6 days.

Lily Pollen Growth in vitro and Agrobacterium-mediated GUS Gene Transformation via Vacuum-Infiltration

  • Park, In-Hae;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.151-154
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    • 2002
  • Conditions for lily pollen growth in vitro and transformation were optimized. Active pollen tube development was achieved effectively in a medium containing 7% sucrose with pH adjusted to 5.7 at the temperature of 27$^{\circ}C$ for about 16-24 hours. Pollen growth was little impaired by the presence of kanamycin at concentration up to 100 mg/L. Pollen rains near the beginning of germination stage were more reliable for Agrobacterium-mediated GUS DNA transformation via vacuum infiltration lasted for 20-40 minutes. GUS DNA integration and its expression in fully developed pollen tubes could be confirmed by Southern blot hybridization, RT-PCR and histochemical staining.

The method to estimate 3-D coordinates of lower trunk muscles using orientation angles during a motion (몸통 운동시 지향각(Orientation angles)을 이용한 허리 근육의 3차원 위치 좌표 추정 기법)

  • Lim, Young-Tae
    • Korean Journal of Applied Biomechanics
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    • v.12 no.1
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    • pp.125-133
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    • 2002
  • The purpose of this study was to develop a method for estimating 3-D coordinates of lower trunk muscles using orientation angles during a motion. Traditional 3-D motion analysis system with DLT technique was used to track down the locations of eight reference markers which were attached on the back of the subject. In order to estimate the orientations of individual lumbar vertebrae and musculoskeletal parameters of the lower trunk muscle, the rotation matrix of the middle trunk reference frame relative to the lower trunk reference frame was determined and the angular locations of individual lumbar vertebrae were estimated by partitioning the orientation angles (Cardan angles) that represent the relative angles between the rotations of the middle and lower trunks. When the orientation angles of individual intervertebral joints were known at a given instant, the instantaneous coordinates of the origin and insertion for all selected muscles relative to the L5 local reference frame were obtained by applying the transformation matrix to the original coordinates which were relative to a local reference frame (S1, L4, L3, L2, or L1) in a rotation sequence about the Z-, X- and Y-axes. The multiplication of transformation matrices was performed to estimate the geometry and kinematics of all selected muscles. The time histories of the 3-D coordinates of the origin and insertion of all selected muscles relative to the center of the L4-L5 motion segment were determined for each trial.

Improved in vitro Regeneration of Potato (Solanum tuberosum cv. Superior) Transformed by Agrobacterium Expressing $\beta-Glucuronidase$

  • Park, Yoon-Kyung;Park, Gene-Sue;Yang, Young-Ki;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.93-98
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    • 1996
  • In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

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LINEAR TRANSFORMATIONS THAT PRESERVE TERM RANK BETWEEN DIFFERENT MATRIX SPACES

  • Song, Seok-Zun;Beasley, Leroy B.
    • Journal of the Korean Mathematical Society
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    • v.50 no.1
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    • pp.127-136
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    • 2013
  • The term rank of a matrix A is the least number of lines (rows or columns) needed to include all the nonzero entries in A. In this paper, we obtain a characterization of linear transformations that preserve term ranks of matrices over antinegative semirings. That is, we show that a linear transformation T from a matrix space into another matrix space over antinegative semirings preserves term rank if and only if T preserves any two term ranks $k$ and $l$.

Studies on the Induction of Transformation in Cereal Plants. III. Cultures and Regeneration of Rice Protoplasts Transferred Foreign Genes. (곡물류의 형질전환 유도에 관한 연구 III. 외래 유전자가 도입된 벼 원형질체의 배양 및 재분화)

  • Hwang, Baik;Hwang, Sung-Jin;Im, Hyong-Tak;Kang, Young-Hee
    • KSBB Journal
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    • v.8 no.1
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    • pp.62-68
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    • 1993
  • Transformed rice plantlet were recovered from protoplasts by electroporation with the plasmld pB 1121, which contain the plant expressible NPT-II and GUS genes. Embryonic cell suspension culture was established with embryonic callus induced from mature seeds of rice (Oryza sativa L. cv. Dong-jin) on the MS medium supplemented with 2.0 mg/l 2,4-D, 0.5 mg/l kinetin, 3% sucrose. Protoplasts isolated from embryonic cell suspensions were electroplated and then poterltialty-transformed tissues were selected by growth on the medium containing 200 mg/l kanamycin sulfate. When subjected to GUS assay, they stained blue, indicating the expression of the inserted GUS genes. Plantlets were regenerated from electroplated protoplasts on the hormone free MS medium. Transferred foreign genes in the plants were confirmed by southern hybridization. These results support use of electroporation for transformation of these important cereal plants.

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An Efficient Plant Regeneration and Transformation System of Robinia pseudoacacia var. umbraculifera for Phytoremediation

  • Kwon, Hye-Jin;Woo, Seong-Min;Seul, Eun-Jun;Kim, Teh-Ryung;Shin, Dong-Un;Kim, Hag-Hyun
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.293-298
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    • 2007
  • Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

Transformation of Populus alba × P. glandulosa by Agrobacterium rhizogenes (Agrobacterium rhizogenes 에 의한 현사시나무의 형질전환(形質轉換))

  • Chung, Kyung Ho;Park, Young Goo;Noh, Eui Rae;Chun, Young Woo
    • Journal of Korean Society of Forest Science
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    • v.78 no.4
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    • pp.372-380
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    • 1989
  • The widely cultivated hybrid poplar Populus alba ${\times}$ P. glandulosa in Korea was transformed with Agrobacterium rhizogenes agropine type strain A4. Genetic transformation was confirmed by the presence of agropine. Alteration in growth rate of hairy roots was seen following changes in the dilution rate of medium and concentration of sucrose, suggesting that improved growth might be achieved by more precise manipulation of the nutrient medium. Plant regeneration occurred from transformed hairy roots on MS medium supplemented with 0.5 mg/l BAP. Transformed plantlets grown in vitro exhibited a more developed root system characterized by fast growth behavior in comparison to normal plantlets. This work demonstrates that the root transformation would be useful in improving plantlet establishment and growth through the effective root system.

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Effects of Wounding and Inoculation Time on Agrobacterium -mediated Transformation in Capsicum annuum L. (상처처리와 접종시간이 Agrobacterium에 의한 고추 형질전환에 미치는 영향)

  • Jeon, Young-Ju;Park, Young-Doo;Choi, Geun-Won
    • Horticultural Science & Technology
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    • v.18 no.6
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    • pp.797-801
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    • 2000
  • The present study was conducted to improve the efficiency of transformation mediated by Agrobacterium tumefaciens in hot pepper. Both regeneration ratio and transformation frequency after the cocultivation with A. tumefaciens were affected by inoculation time and artificial wounding. Transformation frequency was increased over 50% by combining artificial wounding with 120 s of inoculation treatment. Confirmation for the transformation of regenerated shoots was carried out by histochemical ${\beta}$-glucuronidase assay and polymerase chain reaction analysis using npt II primer.

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