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Transfer of SOD2 or NDP kinase 2 genes into purebred lines of petunia

  • Lee, Su-Young;Han, Bong-Hee;Noh, Eun-Woon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.144-148
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    • 2009
  • The transfer of Mn-Superoxide Dismutase (SOD2) gene, complex gene (SA) of CuZnSOD and ascorbate peroxidase (APX), and NDP kinase 2 (NDPK2) gene into Korean 4 cultivars (cvs. Millenium White, Glory Blue, Glory Red, and Glory Purple) and 15 purebred lines of petunia was conducted using Agrobaterium-mediated technique. Two (Wongyo A2-16 and A2-36) of 15 purebred lines and one (cv. Glory Red) of 4 cultivars were effective for the transfer of SOD2 gene. The putative transgenic plants survived on the 2nd selection medium were 124. From PCR analysis, 118 (derived from 4 cultivars and 2 purebred lines) of 124 plants were confirmed to contain marker (npt II ) gene, while 58 of 118 plants did not have target genes. There were no plants with both npt II and SA genes. Twenty seven of 28 SOD2 transgenic plants were re-confirmed as transformants by Sothern analysis. SOD2 and NDPK2 genes were expressed in the transgenic petunias as the ratio of 77.8 to 100.0 % and 23.5%, respectively. T1 seeds were obtained from 36 acclimated transgenic plants (SOD2 34 plus NDPK2) in a glasshouse by self-pollination.

An Effective Method of Diagnosis of Potato Leafroll Virus by RT-PCR (RT-PCR 방법을 이용한 효과적인 감자 잎말림 바이러스의 검정)

  • Jeon, Jae-Heung;Joung, Young-Hee;Choi, Kyung-Hwa;Kim, Hyun-Soon;Oh, Hyun-Woo;Park, Se-Won;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.12 no.3
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    • pp.358-362
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    • 1996
  • 감자 잎말림 바이러스를 검정하기 위하여 ELISA 및 전자현미경에 의해 바이러스 감염이 확인된 기내 배양중인 감장의 줄기로부터 RT-PCR 분석을 수행하였다. 분리된 총 RNA들로부터 바이러스 cDNA를 합성하고 감자 잎말림 바이러스 외피단백질의 일부인 465bp를 특이하게 증폭하도록 고안한 두 primer를 사용하여 PCR 반응을 하였다. 증폭된 465pb의 DNA 절편의 염기서열을 분석한 결과 역시 감자 잎말림 바이러스임을 확인하였다. 바이러스 검정에 있어서 EL-ISA 방법과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법이 ELISA 방법보다 감자 잎말림 바이러스검정에 있어서보다 정확한 방법인 것으로 사료된다.

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Prospects for Plant Biotechnology and Bioindustry in the 21st Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망: 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.19-25
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

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In silico analysis of MeJA-induced comparative transcriptomes in Brassica oleraceae L. var. capitata

  • Lee, Ok Ran;Kim, Dae-Soo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.189-203
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    • 2016
  • Brassica oleraceae var capitata is a member of the Brassicaceae family and is widely used as an horticultural crop. In the present study, transcriptome analysis of B. oleraceae L. var capitata was done for the first time using eight-week old seedlings treated with $50{\mu}m$ MeJA, versus mock-treated samples. The complete transcripts for both samples were obtained using the GS-FLX sequencer. Overall, we obtained 275,570 and 266,457 reads from seedlings treated with or without $50{\mu}m$ MeJA, respectively. All the obtained reads were annotated using biological databases and functionally classified using gene ontology (GO), the Kyoto Encyclopedia of Genes and Genomics (KEGG). By using GO analyses, putative transcripts were examined in terms of biotic and abiotic stresses, cellular component organization, biogenesis, and secondary metabolic processes. The KEGG pathways for most of the transcripts were involved in carbohydrate metabolism, energy metabolism, and secondary metabolite synthesis. In order to double the sequenced data, we randomly chose two putative genes involved in terpene biosynthetic pathways and studied their transcript patterns under MeJA treatment. This study will provide us a platform to further characterize the genes in B. oleracea var capitata.

Mass production and application of activation tagged hairy root lines for functional genomic of secondary metabolism in ginseng

  • Choi, Dong-Woog;Chung, Hwa-Jee;Ko, Suk-Min;In, Dong-Soo;Song, Ji-Sook;Woo, Sung-Sick;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.294-300
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    • 2009
  • Activation tagging that uses T-DNA vectors containing multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an activation tagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosynthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quantitative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of triterpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

Resistance to Potato Virus Y Conferred by PVY Replicase Gene Sequence in Transgenic Burley Tobacco (감자바이러스 Y 복제 유전자로 형질전환된 버어리종 연초의 PVY에 대한 저항성 특성)

  • Young Ho Kim;Eun Kyung Park;Soon Yong Chae;Sang Seock Kim;Kyung-Hee Paek;Hye Sun Cho
    • Journal of the Korean Society of Tobacco Science
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    • v.20 no.1
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    • pp.50-56
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    • 1998
  • The complementary DNA (cDNA) of potato virus Y- vein necrosis strain (PVY-VN) replicase gene (Nlb) was transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Out of 25 putative transformants regenerated, 3 were resistant to PVY-VN, one highly resistant plant with no symptom until seed harvest time and the other two with mild chlorotic spot symptoms at late stages after infection. No symptom was observed in the highly resistant plant, while mild vein necrotic symptoms were developed on suckers of the moderately resistant plants after seed harvest time, In the first generation (T1) via self fertilization, resistance to susceptibility frequency in transgenic plants from the highly resistant transformant was about 3 : 1, while it was lowered much (about 1:2 and 1:19) in T1 of the moderately resistant transformants. In the second generation (T2) of the highly resistant plant, resistance frequencies were similar to T1, but resistance levels varied greatly and appeared to be decreased. Key words : potato virus Y, viral replicate gene, transgenic tobacco plants, resistance.

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Development of KHapmap Browser using DAS for Korean HapMap Research

  • Jin, Hoon;Kim, Seung-Ho;Kim, Young-Uk;Park, Young-Kyu;Ji, Mi-Hyun;Kim, Young-Joo
    • Genomics & Informatics
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    • v.6 no.2
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    • pp.57-63
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    • 2008
  • The Korean HapMap Project has been carried out for the last 5 years since it started in June, 2003. The project generated data for a sum of 1,764,000 Korean SNPs and formally registered the data to the dbSNP of NCBI (The dbSNP website. 2008). We have developed a series of software programs for association studies as well as for the comparison and analysis of Korean HapMap data with four other populations (CEPH, Yoruba, Han Chinese, and Japanese populations). The KHapmap Browser was developed and integrated to provide haplotype retrieval and comparative study tools of human ethnicities for comprehensive disease association studies (http://www.khapmap.org). On that basis, GBrowse was adopted in the KHapmap Browser for inherent Korean genetic data, and a provision of extended services was pledged with the distributed sequence annotation system (DAS). The dynamic linking service of the KHapmap Browser to other tools in our intranetwork environment provides many enhanced functions over GBrowse without DAS. KHapmap Browser is expected to be an invaluable tool for the study of Korean and international Hapmap data.

Histone H4 is cleaved by granzyme A during staurosporine-induced cell death in B-lymphoid Raji cells

  • Lee, Phil Young;Park, Byoung Chul;Chi, Seung Wook;Bae, Kwang-Hee;Kim, Sunhong;Cho, Sayeon;Kang, Seongman;Kim, Jeong-Hoon;Park, Sung Goo
    • BMB Reports
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    • v.49 no.10
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    • pp.560-565
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    • 2016
  • Granzyme A (GzmA) was first identified as a cytotoxic T lymphocyte protease protein with limited tissue expression. A number of cellular proteins are known to be cleaved by GzmA, and its function is to induce apoptosis. Histones H1, H2B, and H3 were identified as GzmA substrates during apoptotic cell death. Here, we demonstrated that histone H4 was cleaved by GzmA during staurosporine-induced cell death; however, in the presence of caspase inhibitors, staurosporine-treated Raji cells underwent necroptosis instead of apoptosis. Furthermore, histone H4 cleavage was blocked by the GzmA inhibitor nafamostat mesylate and by GzmA knockdown using siRNA. These results suggest that histone H4 is a novel substrate for GzmA in staurosporine-induced cells.

Transformations of 2,4,6-Trinitrotoluene in Various Conditions by Klebsiella sp. Strain C1 Isolated from Activated Sludge

  • Chang, Chong-Suk;Kim, Hyoun-Young;Kang, Yang-Mi;Bae, Kyung-Sook;Song, Hong-Gyu
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.193-198
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    • 2002
  • Several 2,4,6-trinitrotoluene (TNT) degrading bacteria were isolated from an activated sludge by an enrichment culture technique, and their TNT removal activities were examined. Among the isolates, strain C1 showed the highest degrading capability, and completely removed 100 or 200 mg I$\^$-1/ of TNT within 6 hours of incubation. This bacterium was identified as Klebsiella sp. The effects of different carbon sources on the removal of the parent TNT by Klebsiella sp. C1 were negligible, but the transformation rates of TNT metabolites such as amino-dinitrotoluenes and diamino-nitrotoluenes were higher with fructose addition compared to glucose addition. When nitrate was used as the nitrogen source, the degradation rates of TNT and hydroxylamino-dinitrotoluenes were higher than those with the ammonium addition. Although the TNT removal rate of Klebsiella sp. C1 was slightly higher in anaerobic conditions, the further transformations of TNT metabolites were more favorable in aerobic conditions.

Characterization of Antimicrobial Substance Producing Lactococcus sp. HM58 Isolated from Gastrointestinal Track of Flounder

  • Jeong Hyun-Mi;Yum Do-Young;Lee Jung-Ki;Choi Mi-Young;Kim Jin-Man
    • Fisheries and Aquatic Sciences
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    • v.6 no.1
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    • pp.27-33
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    • 2003
  • A lactic acid bacterium showing antimicrobial activity against fish pathogen was isolated from gastrointestinal tract of flounder for the purpose of use as an aquaculture probiotics. From the analysis of morphological and physiological characteristics, the isolated strain was named as Lactococcus sp. HM58. Antimicrobial substance (AMS) from Lactococcus sp. HM58 showed strong growth inhibitory activity against Streptococcus sp., which is a fish pathogenic bacterium. AMS was presumed a proteinaceous compound with stability in heat and wide pH range from 2 to 10. It was started to produce in exponential growth phase and was not produced any more in stationary phase. It showed comparatively broad antimicrobial spectrum against most of gram positive bacteria used for this study. About $84\%$ of Lactococcus sp. HM58 was able to survive in the artificial gastric juice though it was low to the extent in the artificial bile juice. In the sensitivity test for various antibiotics, this strain was highly sensitive for doxycycline, erythromycin, amoxicillin clavu1anic acid and ampicillin.