• Title/Summary/Keyword: gene engineering

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Characteristic as a Gene Delivery System of Water Soluble Chitosan Conjugated with Cationic Peptide (양이온 펩타이드가 컨쥬게이트된 수용성 키토산의 유전자 전달체로서의 특성)

  • Kim, Young-Min;Kim, Ji-Ho;Park, Seong-Cheol;Park, Yung-Hoon;Jang, Mi-Kyeong
    • KSBB Journal
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    • v.31 no.4
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    • pp.300-311
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    • 2016
  • Recently gene delivery has been designed newly using bioactive biomaterial and applied in the various field by many researchers. In this study, we proposed a new gene delivery system which has the capability of targeting effect in the specific tissue and remarkably enhanced transfection efficiency. We investigated $^1H-NMR$ spectroscopy, particle size analyzer and gel retardation to confirm the correct preparation of gene delivery. Also, we identified the hemo-compatibility of gene delivery by hemolysis assay, non-cytotoxicity by MTT test and transfection efficiency. The uptake mechanism of the gene carrier was confirmed using inhibitor agent such as sodium azide, indomethacin, quercetin, colchicine, and chloropromazine. As a results, it was identified that gene carrier prepared by in this study entered in the cell by the microtubule-dependent, energy-dependent and clathrin-mediated endocytosis pathway.

Parthenolide-Induced Apoptosis, Autophagy and Suppression of Proliferation in HepG2 Cells

  • Sun, Jing;Zhang, Chan;Bao, Yong-Li;Wu, Yin;Chen, Zhong-Liang;Yu, Chun-Lei;Huang, Yan-Xin;Sun, Ying;Zheng, Li-Hua;Wang, Xue;Li, Yu-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4897-4902
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    • 2014
  • Purpose: To investigate the anticancer effects and underlying mechanisms of parthenolide on HepG2 human hepatocellular carcinoma cells. Materials and Methods: Cell viability was assessed by MTT assay and cell apoptosis through DAPI, TUNEL staining and Western blotting. Monodansylcadaverin(MDC) and AO staining were used to detect cell autophagy. Cell proliferation was assessed by Ki67 immunofluorescence staining. Results: Parthenolide induced growth inhibition in HepG2 cells. DAPI and TUNEL staining showed that parthenolide could increase the number of apoptotic nuclei, while reducing the expression of the anti-apoptotic protein Bcl-2 and elevating the expression of related proteins, like p53, Bax, cleaved caspase9 and cleaved caspase3. Parthenolide could induce autophagy in HepG2 cells and inhibited the expression of proliferation-related gene, Ki-67. Conclusions: Parthenolide can exert anti-cancer effects by inducing cell apoptosis, activating autophagy and inhibiting cell proliferation.

Molecular Characterization and Expression Patterns of Porcine Eukaryotic Elongation Factor 1 A

  • Wang, H.L.;Wang, H.;Zhu, Z.M.;Yang, S.L.;Fen, S.T.;Li, Kui
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.953-957
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    • 2006
  • The eukaryotic elongation factor 1 A (EEF1A) participates in protein synthesis by forming the eEF1A GTP tRNA complex to deliver aminoacyl-tRNA to the A site of ribosomes. This study described cDNA sequences and partial genomic structure of porcine EEF1A1. The porcine EEF1A1 gene encoded a protein with 462 amino acids, which shared complete homology with human, chimpanzee and dog. The temporal expression pattern showed the diversity of EEF1A1 level in mRNA was relatively minor in prenatal embryo skeletal muscle, however, the expression decreased during aging after birth in skeletal muscle of the Chinese Tongcheng pig. The spatial expression patterns indicated that the gene expressed in skeletal muscle, heart, lung, liver, kidney, fat and spleen. In addition, we assigned the gene to porcine chromosome 1 using a radiation hybrid panel.

Studies on QTLs for Bakanae Disease Resistance with Populations Derived from Crosses between Korean japonica Rice Varieties

  • Dong-Kyung Yoon;Chaewon Lee;Kyeong-Seong Cheon;Yunji Shin;Hyoja Oh;Jeongho Baek;Song-Lim Kim;Young-Soon Cha;Kyung-Hwan Kim;Hyeonso Ji
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.201-201
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    • 2022
  • Rice bakanae disease is a serious global threat in major rice-cultivating regions worldwide causing high yield loss. It is caused by the fungal pathogen Fusarium fujikuroi. Varying degree of resistance or susceptibility to bakanae disease had been reported among Korean japonica rice varieties. We developed a modified in vitro bakanae disease bioassay method and tested 31 Korean japonica rice varieties. Nampyeong and Samgwang varieties showed highest resistance while 14 varieties including Junam and Hopum were highly susceptible with 100% mortality rate. We carried out mapping QTLs for bakanae disease resistance with four F2:F3 populations derived from the crosses between Korean japonica rice varieties. The Kompetitive Allele-Specific PCR (KASP) markers developed in our laboratory based on the SNPs detected in Korean japonica rice varieties were used in genotyping F2 plants in the populations. We found four major QTLs on chromosome 1, 4, 6, and 9 with LOD scores of 21.4, 6.9, 6.0, and 60.3, respectively. In addition, we are doing map-based cloning of the QTLs on chromosome 1 and 9 which were found with Junam/Nampyeong F2:F3 population and Junam/Samgwang F2:F3 population, respectively. These QTLs will be very useful in developing bakanae disease resistant high quality rice varieties.

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PCR-mediated Recombination of the Amplification Products of the Hibiscus tiliaceus Cytosolic Glyceraldehyde-3-phosphate Dehydrogenase Gene

  • Wu, Linghui;Tang, Tian;Zhou, Renchao;Shi, Suhua
    • BMB Reports
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    • v.40 no.2
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    • pp.172-179
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    • 2007
  • PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

Gene Selection using Principal Component Analysis for Molecular classification (Principal Component Analysis를 이용한 Gene Selection)

  • Lim Soo-Hong;Sohn Kirack;Hong Sung-Yong
    • Proceedings of the Korean Information Science Society Conference
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    • 2005.07b
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    • pp.259-261
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    • 2005
  • 수천개의 Gene Expression Measurement를 생성해 내는 DNA Microarray 연구는 조직과 세포의 표본으로부터 진단에 유용한 Gene Expression 정보를 모으게 된다. 이런 종류의 Data를 분석하기 위하여 SVM(Support Vector Machine)을 사용한 새로운 방법이 연구되어왔다. 본 논문에서는 Gene Expression Data에 대한 고유벡터(Eigen Vector)를 이용하여 SVM의 성능을 향상시키고 질병진단에 유용한 Gene을 찾아 내는 알고리즘을 기술한다. 고유벡터를 통하여 Gene을 선택적으로 SVM Learning에 참가 시키고 분류의 결과를 통하여 추가된 Gene이 질병 진단에 미치는 영향력을 알아냄으로써 질병에 대한 Gene 역할을 파악 하는데 활용할 수 있다.

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Functional characterization of ABA signaling components using transient gene expression in rice protoplasts

  • Song, In-Sik;Moon, Seok-Jun;Kim, Jin-Ae;Yoon, Insun;Kwon, Taek-Ryoun;Kim, Beom-Gi
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.109-109
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    • 2017
  • The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through interaction networks resulting in diverse signaling responses. We tried to characterize those gene functions using transient gene expression for rice protoplasts (TGERP) because it is instantaneous and convenient system. Firstly, in order to monitor the ABA signaling output, we developed reporter system named pRab16A-fLUC which consists of Rab16A promoter of rice and luciferase gene. It responses more rapidly and sensitively to ABA than pABRC3-fLUC that consists of ABRC3 of HVA1 promoter in TGERP. We screened the reporter responses for over-expression of each signaling components from group A OsbZIPs to OsPYL/RCARs with or without ABA in TGERP. OsbZIP46 induced reporter most strongly among OsbZIPs tested in the presence of ABA. SAPKs could activate the OsbZIP46 even in the ABA independence. Subclass A OsPP2C6 and -8 almost completely inhibited the OsbZIP46 activity in the different degree through the SAPK9. Lastly, OsPYL/RCAR2 and -5 rescued the OsbZIP46 activity in the presence of SAPK9 and OsPP2C6 dependent on ABA concentration and expression level. By using TGERP, we could characterize successfully the effects of ABA dependent gene expression signaling components in rice. In conclusion, TGERP represents very useful technology to study systemic functional genomics in rice or other monocots.

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Cloning and Characterization of a PI-like MADS-Box Gene in Phalaenopsis Orchid

  • Guo, Bin;Hexige, Saiyin;Zhang, Tian;Pittman, Jon K.;Chen, Donghong;Ming, Feng
    • BMB Reports
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    • v.40 no.6
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    • pp.845-852
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    • 2007
  • The highly evolved flowers of orchids have colorful sepals and fused columns that offer an opportunity to discover new genes involved in floral development in monocotyledon species. In this investigation, we cloned and characterized the homologous PISTALLATA-like (PI-like) gene PhPI15 ($\underline{Ph}alaenopsis$ $\underline{PI}$ STILLATA # $\underline{15}$), from the Phalaenopsis hybrid cultivar. The protein sequence encoded by PhPI15 contains a typical PI-motif. Its sequence also formed a subclade with other monocot PI-type genes in phylogenetic analysis. Southern analysis showed that PhPI15 was present in the Phalaenopsis orchid genome as a single copy. Furthermore, it was expressed in all the whorls of the Phalaenopsis flower, while no expression was detected in vegetative organs. The flowers of transgenic tobacco plants ectopically expressing PhPI15 showed male-sterile phenotypes. Thus, as a Class-B MADS-box gene, PhPI15 specifies floral organ identity in orchids.

Chemical Synthesis and Cloning of Panax ginseng Peptide Gene

  • Zhang, Hong-Ying;Chen, Dong-Song;Zhang, Jin
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.65-67
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    • 1990
  • The sequence of ginseng peptide gene was designed and synthesized by the solid phase plasmid pUC19. Escherichia coli JM101 cells were transformed with above hybrid plasmids. Ampicillin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techinques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

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Reverse Engineering of a Gene Regulatory Network from Time-Series Data Using Mutual Information

  • Barman, Shohag;Kwon, Yung-Keun
    • Proceedings of the Korea Information Processing Society Conference
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    • 2014.11a
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    • pp.849-852
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    • 2014
  • Reverse engineering of gene regulatory network is a challenging task in computational biology. To detect a regulatory relationship among genes from time series data is called reverse engineering. Reverse engineering helps to discover the architecture of the underlying gene regulatory network. Besides, it insights into the disease process, biological process and drug discovery. There are many statistical approaches available for reverse engineering of gene regulatory network. In our paper, we propose pairwise mutual information for the reverse engineering of a gene regulatory network from time series data. Firstly, we create random boolean networks by the well-known $Erd{\ddot{o}}s-R{\acute{e}}nyi$ model. Secondly, we generate artificial time series data from that network. Then, we calculate pairwise mutual information for predicting the network. We implement of our system on java platform. To visualize the random boolean network graphically we use cytoscape plugins 2.8.0.