• Title/Summary/Keyword: K-L transformation

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A Novel Mutant of Human Papillomavirus Type 18 E6E7 Fusion Gene and its Transforming Activity

  • Zhou, Zhi-Xiang;Zhao, Chen;Li, Qian-Qian;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7395-7399
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    • 2014
  • Background: Persistent human papillomavirus (HPV) infection, especially with high-risk types such as HPV16 and HPV18, has been identified as the primary cause of cervical cancer. E6 and E7 are the major onco-proteins of high-risk HPVs, which are consistently expressed in HPV infected tissues but absent in normal tissues and represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this study, the optimized fusion gene HPV18 E6E7 (HPV18 ofE6E7) was constructed according to genetic codon usage for human genes. At the same time, for safety future clinical application, a mutant of HPV18 ofE6E7 fusion gene was generated by site-directed mutagenesis at L52G for the E6 protein and C98G for the E7 protein. Results: HPV18-E6E7 mutant (HPV18 ofmE6E7) constructed in this work not only lost the transformation capability for NIH 3T3 cells and tumorigenicity in BALB/c nude mice, but also maintained very good stability and antigenicity. Conclusion: These results suggest that the mutant should undergo further study for application as a safe antigenspecific therapeutic vaccine for HPV18-associated tumors.

Production of Transgenic Birdsfoot trefoil Plants by Introduction of 'SWPA2 Promoter + AtNDPK2 Gene' ('SWPA2프로모터+AtNDPK2유전자' 도입에 의한 버즈풋 트레포일 형질전환체 생산)

  • Kim Ki-Yong;Jang Yo-Soon;Kim Meing Jooung;Lim Keun Bal;Kim Won Ho;Seo Sung;Lee Sang Jin;Kwak Sang-Soo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.4
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    • pp.281-286
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    • 2005
  • To develop transgenic birdsfoot trefoil (Lotus corniculatus L.) plants tolerant to environmental stress, Arabidopsis NDPK gene (AtNDPK) was introduced into birdsfoot trefoil plants using Agrobacterium-mediated transformation and expressed powerfully under the control of the SWPA2 promoter. The expression vector, pCAMBIA2300 was used for introduction of AtNDPK gene into birdsfoot trefoil plants. The transformed calli were selected on kanamycin containing medium and then regenerated. The transformed birdsfoot trefoil plants were cultivated fur 4 months on BOi2Y medium. Genomic DNA PCR and Southern blot analysis confirmed the incorporation of AtNDPK into the birdsfoot trefoil genome.

The Protective Effects of EGCG Extracted from Green Tea on Apoptosis Induced by$H_2O_2$ in Conjunctival Cell Lines ($H_2O_2$에 의한 결막 세포주의 세포고사에서 녹차추출물 EGCG의 보호효과)

  • Park, Su-Kyoung;Chae, Soo-Chul;Kho, Eun-Gyeong;Ryu, Geun-Chang;Kim, Jai-Min;Na, Myung-Suk;Lee, Jong-Bin
    • Journal of Korean Ophthalmic Optics Society
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    • v.13 no.3
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    • pp.95-101
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    • 2008
  • Purpose: Hydrogen peroxide which is one of the reactive oxygen species has been seen to cause various diseases, various cellular disinfections, gene transformation and cell death. The goals of this study were to determine the protective effect of EGCG against $H_2O_2$-induced apoptotic death in conjunctival cell lines. Methods: We measured cell viability by MTT assay and analyzed DNA fragmentation to check up a distinctive feature in cell death and measured the removal ability of free radicals by DPPH free radical scavenging assay and evaluated the oxygen free radical's quantity in the cell by DCFH-DA assay. The mRNA expression in the cell were examined by RT-PCR. Results: Cell viability and free radical scavening activites was significantly increased in dose dependently after cell was exposed to EGCG. And DNA fragmentation and intracellular ROS was decreased. It was showed the mRNA expression which increase of bcl-2, bcl-xL expression and decrease of bax expression. Conclusions: From these results, it suggests that EGCG has an antioxidant effect and protects conjunctival cell lines from the $H_2O_2$-mediated apoptosis through the modulation of the mRNA expression.

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Increment of fructan biosynthesis in rice by transformation of 1-sst and 1-fft genes isolated from jerusalem artichoke (Helianthus tuberosus L.) (돼지감자 유래 1-sst와 1-fft 유전자의 형질전환 발현에 의한 벼의 fructan 생합성 증진)

  • Kang, Kwon-Kyoo;Song, Beom-Heon;Lee, Gyong-A;Lee, Hye-Jung;Park, Jin-Ha;Jung, Yu-Jin;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.102-109
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    • 2010
  • Fructan has been found to accumulate in various tissues during periods when light levels increased carbon fixation where low temperatures reduced growth rates while photosynthesis continued. In this study, we have cloned 1-sucrose:sucrose fructosyl transferase(1-sst) and 1-fructan: fructan fructosyl transferase (1-fft, a key enzyme for the synthesis of fuctan) from Jerusalem Artichoke (Helianthus tuberosus L.). The recombinant vector with 1-sst and 1-fft has been constructed under the control of 35S promoter of KJGV-B2 vector and transgenic plants obtained by Agrobacterium tumefaciens LBA4404. PCR analysis carried out on the putative transgenic plants for amplification of the coding region of specific gene (1-sst, 1-fft), and HPT genes. Transgenic lines carrying of 1-sst and 1-fft were confirmed for integration into the rice genome using Southern blot hybridization and RT-PCR. The transgenic plants in $T_2$ generation were selected and expression pattern analysis revealed that 1-sst and 1-fft were stable. This analysis confirmed the presence of low-molecular-weight fructan in the seedling of the transgenic rices. Therefore, cold tolerance and carbohydrate metabolism will be possible to develop resistant plants using the transgenic rice.

Isolation of Superoxide Dismutase cDNAS from an Weedy Rice Variety and Transformation of a Cultivated Rice Variety (잡초성벼의 superoxide dismutase cDNA cloning과 재배벼로의 형질전환)

  • Park, Sang-Gyu;Park, Jong-Suk;Lee, Seung-In;Suh, Suk-Chul;Kim, Byung-Keuk;Jo, Youl-Lae;Suh, Hak-Soo
    • Korean Journal of Environmental Agriculture
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    • v.21 no.2
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    • pp.156-161
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    • 2002
  • Two different cDNA clones for superoxide dismutase (SOD) were isolated from an weedy rice variety (Oryza sativa, cv. Bhutan14Ad) and were introduced into a cultivated rice variety (Oryza sativa, cv. Nakdong) in order to develop the environmental stress-resistant rice plants. Sequence analysis of the cloned cDNAS indicated that the deduced amino acid sequence of SOD-A is 88.4% identical to that of SOD-B. Furthermore, the nucleotide sequence of SOD-A is 99.3% identical to that of a Cu/Zn SOD gene of Oryza sativa (GenBank accession No. L36320). The nueleotide sequence of SOD-B was identical to that of the previously published SOD gene (Accession No. D01000). A cultivated rice variety, Nakdong-byeo, was transformed with chimeric SOD genes containing a actin promoter of rice and pin2 terminator using a particle bombardment technique. Transformed calli were selected on an selection medium containing phosphinothricin (PPT). Transgenic rice plants were regenerated from the PPT-resistant calli. PCR analysis with genomic DNAs from transgenic plants revealed that transgenes are introduced into rice genome.

Production Yield Enhancement of Mycosporine-like amino acid(MAA)s in Transformed Microalgae Culture by Radiofrequency (형질전환 미세조류의 고주파 처리 배양을 통한 MAA 생산량 증가)

  • Seo, Hyo Hyun;Song, Mi Young;Kulkarni, Atul;Suh, Sung-Suk;Lee, Taek-Kyun;Moh, Sang Hyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.6
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    • pp.3799-3804
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    • 2014
  • In sea water, microalgae are exposed to a range of critical environmental conditions. Microalgae are protected from UV-A radiation due to the presence of mycosporine like amino acids(MAAs). Owing to the UV-A absorption properties of MAAs, they are used widely as a UV protecting ingredient in cosmetics. Therefore, there is a need to increase the production yield of MAAs. This study investigated the production yield of MAAs in transformed microalgae by radiofrequency(RF) exposure. Initially, the Glut-1 gene was transformed to Chlamydomonas hedleyi microalgae as a glucose transporter. The biomass was enhanced after Glut-1 gene transformation. In addition, the MAAs production yield was increased during large scale production in bioreactors due to the RF treatment. Therefore, purified extracts of MAAs can be used as a sun block material in the cosmetic industrial field.

Development of transgenic cucumbers expressing Arabidopsis Nit gene (애기장대 Nit유전자 발현 오이 형질전환체 개발)

  • Jang, Hyun A;Lim, Ka Min;Kim, Hyun A;Park, Yeon-Il;Kwon, Suk Yoon;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.198-202
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    • 2013
  • To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.

Transgenic Rice Expressing Snowdrop Lectin (Galanthus nivalis agglutinin; GNA) Shows Resistance to Rice Brown Planthopper (Nilaparvata lugens Stål) (Snowdrop lectin (Galanthus nivalis agglutinin: GNA) 유전자 도입에 의한 벼멸구 저항성 형질전환 벼 개발)

  • Lee, Soo In;Yoon, In Sun;Kim, Jin A;Hong, Joon Ki;Park, Beom-Seok;Lee, Yeon-Hee
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1614-1620
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    • 2012
  • Transgenic rice plants with increased resistance to rice brown planthopper (Nilaparvata lugens St${\aa}$l) were generated by particle bombardment-mediated transformation of plants with a gene encoding snowdrop lectin (Galanthus nivalis agglutinin; GNA) under control of the rice Rubisco small subunit (rbcS) promoter.. A large number of transgenic rice plants containing the GNA gene were generated. The integration, expression, and inheritance of this gene in the $R_1$ and $R_2$ generations were demonstrated by Southern and western blot analyses. The plants contained one to five copies of the transgene. The GNA protein comprised approximately 0.01-2.0% of total soluble protein in the $R_1$ and $R_2$ transgenic plants. Insect bioassays and feeding studies showed that the GNA protein expressed in the $R_2$ transgenic rice plants reduced the survival of brown planthoppers. The introduction of GNA into rice plants therefore can help to control insect pests.

A Finite Element Based PML Method for Time-domain Electromagnetic Wave Propagation Analysis (시간영역 전자기파 전파해석을 위한 유한요소기반 PML 기법)

  • Yi, Sang-Ri;Kim, Boyoung;Kang, Jun Won
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.28 no.2
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    • pp.123-130
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    • 2015
  • This paper presents a new formulation for transient simulations of microwave propagation in heterogeneous unbounded domains. In particular, perfectly-matched-layers(PMLs) are introduced to allow for wave absorption at artificial boundaries used to truncate the infinite extent of the physical domains. The development of the electromagnetic PML targets the application to engineering mechanics problems such as structural health monitoring and inverse medium problems. To formulate the PML for plane electromagnetic waves, a complex coordinate transformation is introduced to Maxwell's equations in the frequency-domain. Then the PML-endowed partial differential equations(PDEs) for transient electromagnetic waves are recovered by the application of the inverse Fourier transform to the frequency-domain equations. A mixed finite element method is employed to solve the time-domain PDEs for electric and magnetic fields in the PML-truncated domain. Numerical results are presented for plane microwaves propagating through concrete structures, and the accuracy of solutions is investigated by a series of error analyses.

The Identification of Japanese Black Cattle by Their Faces

  • Kim, Hyeon T.;Ikeda, Y.;Choi, Hong L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.868-872
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    • 2005
  • Individual management of the animal is the first step towards reaching the goal of precision livestock farming that aids animal welfare. Accurate recognition of each individual animal is important for precise management. Electronic identification of cattle, usually referred to as RFID (Radio Frequency Identification), has many advantages for farm management. In practice, however, RFID implementations can cause several problems. Reading speed and distance must be optimized for specific applications. Image processing is more effective than RFID for the development of precision farming system in livestock. Therefore, the aim of this paper is to attempt the identification of cattle by using image processing. The majority of the research on the identification of cattle by using image processing has been for the black-and-white patterns of the Holstein. But, native Japanese and Korean cattle do not have a consistent pattern on the body, so that identification by pattern is impossible. This research aims to identify to Japanese black cattle, which does not have a black-white pattern on the body, by using image processing and a neural network algorithm. 12 Japanese black cattle were tested. Values of input parameter were calculated by using the face image values of 12 cows. The face was identified by the associate neural memory algorithm, and the algorithm was verified by the transformed face image, for example, of brightness, distortion, noise and angle. As a result, there was difference due to a transformation ratio of the brightness, distortion, noise, and angle. The algorithm could identify 100% in the range from -30 to +30 degrees of brightness, -20 to +40 degrees of distortion, 0 to 60% of noise and -20 to +30 degree of angle transformed images.