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Transfection and Expression of Reconstructed Genes within Baculoviral Vectors (Baculovirus 벡터내 재구성된 유전자의 전이와 발현)

  • Sa, Young-Hee;Choi, hang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.05a
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    • pp.588-591
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    • 2018
  • Baculovirus was originally isolated from the alfalfa looper and contains a 134-kbp genome with 154 open reading frames (ORF). The major capsid protein VP39 together with some minor proteins forms the nucleocapsid ($21nm{\times}260nm$) that encloses the DNA with p6.9 protein. They are double-stranded, circular, supercoiled DNA molecules in a rod-shaped capsid. Wild-type baculoviruses exhibit both lytic and occluded life cycles that develop independently throughout the three phases of virus replication. Recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. Especially, inclusion of a dominant selectable marker in these baculoviral vectors can express diverse recombinant genes in many cells. Baculoviral vectors were reconstructed with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) gene and so on. These reconstructed vectors were infected into various cell and cell lines. We performed transfection and expression of these recombinant vectors comparison with other control vectors. From this study, we knew that transfection and expression of these recombinant vectors have higher efficacy than any control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

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Gene Structure of Cotesia plutellae Bracovirus (CpBV)-$I_{k}B$ and Its Expression Pattern in the Parasitized Diamondback Moth, Plutella xylostella (프루텔고치벌 브라코바이러스(Cotesia plutellae Bracovirus) 유래 $I_{k}B$ 유전자 구조와 피기생 배추좀나방(Plutella xylostella) 체내 발현 패턴)

  • Kim Yong-Gyun;Basio Neil A.;Ibrahim Ahmed M.A.;Bae Sung-Woo
    • Korean journal of applied entomology
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    • v.45 no.1 s.142
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    • pp.15-24
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    • 2006
  • Inhibitor kB (IkB)-like gene has been found in the genome of Cotesia plutellae bracovirus (CpBV), which is the obligatory symbiont of an endoparsitoid wasp, C. plutellae. The open reading frame of CpBV-IkB was 417 bp and encoded 138 amino acids. Four ankyrin repeat domains were found in CpBV-IkB, which shared high homology with other known polydnavirus IkBs. Considering a presumptive cellular IkB based on Drosophila Cactus, CpBV-IkB exhibited a truncated structure with deletion of signal-receiving domains, which suggested its irreversible inhibitory role in NFkB signal transduction pathway of the parasitized host in response to the wasp parasitization. CpBV-IkB was expressed only in the parasitized diamondback moth, Plutella flostella. Its expression was estimated by quantitative RT-PCR during parasitization period, showing a constitutive expression pattern from the first day of parasitization. An indirect functional analysis of CpBV-IkB was conducted and suggested a hypothesis of host antivirus inhibition.

Development of Workbench for Analysis and Visualization of Whole Genome Sequence (전유전체(Whole gerlome) 서열 분석과 가시화를 위한 워크벤치 개발)

  • Choe, Jeong-Hyeon;Jin, Hui-Jeong;Kim, Cheol-Min;Jang, Cheol-Hun;Jo, Hwan-Gyu
    • The KIPS Transactions:PartA
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    • v.9A no.3
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    • pp.387-398
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    • 2002
  • As whole genome sequences of many organisms have been revealed by small-scale genome projects, the intensive research on individual genes and their functions has been performed. However on-memory algorithms are inefficient to analysis of whole genome sequences, since the size of individual whole genome is from several million base pairs to hundreds billion base pairs. In order to effectively manipulate the huge sequence data, it is necessary to use the indexed data structure for external memory. In this paper, we introduce a workbench system for analysis and visualization of whole genome sequence using string B-tree that is suitable for analysis of huge data. This system consists of two parts : analysis query part and visualization part. Query system supports various transactions such as sequence search, k-occurrence, and k-mer analysis. Visualization system helps biological scientist to easily understand whole structure and specificity by many kinds of visualization such as whole genome sequence, annotation, CGR (Chaos Game Representation), k-mer, and RWP (Random Walk Plot). One can find the relations among organisms, predict the genes in a genome, and research on the function of junk DNA using our workbench.

Analysis of copy number abnormality (CNA) and loss of heterozygosity (LOH) in the whole genome using single nucleotide polymorphism (SNP) genotyping arrays in tongue squamous cell carcinoma (설편평상피암에 있어서의 고밀도 SNP Genotyping 어레이를 이용한 전게놈북제수와 헤테로접합성 소실의 분석)

  • Kuroiwa, Tsukasa;Yamamoto, Nobuharu;Onda, Takeshi;Bessyo, Hiroki;Yakushiji, Takashi;Katakura, Akira;Takano, Nobuo;Shibahara, Takahiko
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.6
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    • pp.550-555
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    • 2011
  • Chromosomal loss of heterozygosity (LOH) is a common mechanism for the inactivation of tumor suppressor genes in human epithelial cancers. LOH patterns can be generated through allelotyping using polymorphic microsatellite markers; however, owing to the limited number of available microsatellite markers and the requirement for large amounts of DNA, only a modest number of microsatellite markers can be screened. Hybridization to single nucleotide polymorphism (SNP) arrays using Affymetarix GeneChip Mapping 10 K 2.0 Array is an efficient method to detect genome-wide cancer LOH. We determined the presence of LOH in oral SCCs using these arrays. DNA was extracted from tissue samples obtained from 10 patients with tongue SCCs who presented at the Hospital of Tokyo Dental College. We examined the presence of LOH in 3 of the 10 patients using these arrays. At the locus that had LOH, we examined the presence of LOH using microsatellite markers. LOH analysis using Affymetarix GeneChip Mapping 10K Array showed LOH in all patients at the 1q31.1. The LOH regions were detected and demarcated by the copy number 1 with the series of three SNP probes. LOH analysis of 1q31.1 using microsatellite markers (D1S1189, D1S2151, D1S2595) showed LOH in all 10 patients (100). Our data may suggest that a putative tumor suppressor gene is located at the 1q31.1 region. Inactivation of such a gene may play a role in tongue tumorigenesis.

Trends of commercial strain development and spawn industry in mushrooms (버섯의 품종 육성과 종균 산업의 동향)

  • Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Oh, Se-Jong;Cheong, Jong-Chun;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.4 no.1
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    • pp.1-32
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    • 2006
  • According to UPOV (International Union for Protection of New Varieties of Plants), mushroom spawn market have to be opened by the year 2009. Number of commercial strains distributed by the year 2005 were 179 of 24 species of edible and medicinal mushrooms. Only nine strains of oyster mushrooms were registered as protected variety, which is not compatible with those recorded in other advanced countries. Meaning of spawn in broad sense contains commercial strains. Development of commercial strains faces two main problems in Korea : One is the complicated genetic patterns and sexuality of mushroom species, and the other is expensive experimental equipments and fruiting body growing houses. Resolution of these problems leads to development of mushroom strains. This could be achieved as follows; genetic resources collection and assessment, molecular characterization of useful genetic characters, development of new commercial strains by hybridization using typical genetic resources, strengthening of breeding research using "Mushroom Breeding Group", management of spawn research company by consortium, foundation of mushroom general industry, promotion of consumption, and upgrade of competition ability for other countries. These points are under discussion.

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Functional characterization of a CCCH type zinc-finger protein gene OsZF2 by ectopic overexpression of the gene in rice (과발현 형질전환벼에서 CCCH type zinc-finger protein 유전자 OsZF2 기능 분석)

  • Lee, Jung-Sook;Yoon, In-Sun;Yoon, Ung-Han;Lee, Gang-Seob;Byun, Myung-Ok;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.23-29
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    • 2009
  • We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

Selection of transgenic sweetpotato plants expressing 2-Cys peroxiredoxin with enhanced tolerance to oxidative stress (Peroxiredoxin 유전자 발현 산화스트레스 내성 형질전환 고구마의 선발)

  • Kim, Myoung-Duck;Yang, Kyoung-Sil;Kwon, Suk-Yoon;Lee, Sang-Yeol;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.75-80
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    • 2009
  • In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at $20{\mu}M$, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.

Identification and Molecular Characterization of Methionine Sulfoxide Reductase B Gene in Rice Blast Fungus, Magnaporthe oryzae (벼도열병균에서의 methionine sulfoxide reductase B 유전자의 분자적 특성)

  • Kim, Jeong-Hwan;Kim, Jin-Soo;Jeong, Mi-Yeon;Choi, Woo-Bong
    • Journal of Life Science
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    • v.19 no.3
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    • pp.343-348
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    • 2009
  • Magnaporthe oryzae, a major cause of rice blast, is one of the most destructive plant fungal pathogens. Secretion of reactive oxygen species (ROS) during the infection phase of plant pathogenic fungus plays a key role in the defense mechanism of a plant. ROS causes oxidative damage and functional modification to the proteins in a pathogenic fungus. Methionine, especially, is a major target of ROS, which oxidizes it to methionine sulfoxide. To survive from the attack of ROS, plant pathogenic fungus has antioxidative systems - one example would be methionine sulfoxide reductase B (MSRB), which reverses the oxidative alteration of methionine to methionine sulfoxide. In the present study, identification and molecular characterization of the MSRB gene in M. oryzae KJ201 were investigated. The MSRB gene was amplified by PCR from the M. oryzae KJ201 genomic DNA. The copy number of MSRB in the genome of M. oryzae KJ201 was identified by Southern blot analysis, which revealed that the gene exists as a single copy. To study the molecular function of an MSRB gene, the expression level of the MSRB gene was assayed with hydrogen peroxide treatment by Northern blot analysis and RT-PCR. The expression of the MSRB gene was increased by treatment of hydrogen peroxide, without significant correlation to hydrogen peroxide concentrations. These results indicate that the MSRB gene in M. oryzae KJ201 could contribute to protection against plant defense compounds such as ROS and offer a novel strategy for the control of rice blast.

Conservative Genes of Less Orthologous Prokaryotes (Orthologs 수가 적은 원핵생물들의 보존적 유전자)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.27 no.6
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    • pp.694-701
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    • 2017
  • Mycoplasma genitalium represents the smallest genome among mono-cultivable prokaryotes. To discover and compare the orthologs (conservative genes) among M. genitalium and 14 prokaryotes that are uncultivable and have less orthologs than M. genitalium, COG (clusters of orthologous groups of protein) analyses were applied. The analyzed prokaryotes were M. genitalium, one hyperthermophilic exosymbiotic archaeon Nanoarchaeum equitans, four intracellular plant pathogenic eubacteria of Candidatus Phytoplasma genus, and nine endosymbiotic eubacteria of phloem- and xylem-feeding insects. Among 367 orthologs of M. genitalium, 284 orthologs were conservative between M. genitalium and at least one other prokaryote. All 15 prokaryotes commonly have 29 orthologs, representing the significance of proteins in life. They belong to 25 translation-related, including 22 ribosomal proteins, 3 subunits of RNA polymerase, and 1 protein-folding-related. Among the 15 prokaryotes, 40 orthologs were only found in all four Candidatus Phytoplasma. The other nine Candidatus, all endosymbionts with insects, showed only a single common COG0539 (ribosomal protein S1), representing the diversity of orthologs among them. These results might provide clues to understand conservative genes in uncultivable prokaryotes, and may be helpful in industrial areas, such as handling prokaryotes producing amino acids and antibiotics, and as precursors of organic synthesis.

Genetic Analysis Strategies for Improving Race Performance of Thoroughbred Racehorse and Jeju Horse (서러브레드 경주마와 제주마의 경주 능력 향상을 위한 유전체 분석 전략)

  • Baek, Kyung-Wan;Gim, Jeong-An;Park, Jung-Jun
    • Journal of Life Science
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    • v.28 no.1
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    • pp.130-139
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    • 2018
  • In ancient times, horse racing was done in ancient European countries in the form of wagon races or mountain races, and wagon racing was adopted as a regular event at the Greek Olympic Games. Thoroughbred horse has been bred since 17th century by intensive selective breeding for its speed, stamina, and racing ability. Then, in the 18th century, horse racing using the Thoroughbred species began to gain popularity among nobles. Since then, horse racing has developed into various forms in various countries and have developed into flat racing, steeplechasing, and harness racing. Thoroughbred racehorse has excellent racing abilities because of powerful selection breeding strategy for 300 years. It is necessary to maintain and maximize horses' ability to race, because horse industries produce enormous economic benefits through breeding, training, and horse racing. Next-generation sequencing (NGS) methods which process large amounts of genomic data have been developed recently. Based on the remarkable development of these genomic analytical techniques, it is now possible to easily carry out animal breeding strategies with superior traits. In order to select breeding racehorse with superior racing traits, the latest genomic analysis techniques have to be introduced. In this paper, we will review the current efforts to improve race performance for racehorses and to examine the research trends of genomic analysis. Finally, we suggest to utilize genomic analysis in Thoroughbred racehorse and Jeju horse, and propose a strategy for selective breeding for Jeju horse, which contributes job creation of Korea.