• Journal of Microbiology and Biotechnology
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• 제21권4호
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• pp.374-378
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• 2011

To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1894-1901
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• 2015

A novel sulfatase gene, ary423 (1,536 bp ORF), encoding a protein of 511 amino acids with a calculated molecular mass of 56 kDa, was identified from Flammeovirga pacifica, which was isolated from deep-sea sediments of west Pacific Ocean. Amino acid sequence analysis revealed that Ary423 possessed a conserved C-X-A-X-R motif, which was recognized as the sulfatase signature. Phylogenetic analysis suggested that Ary423 belonged to arylsulfatases. After heterologous expression in Escherichia coli cells, the recombinant Ary423 was purified with a Ni⁺ affinity column, and was shown to be highly active at a broad range of temperatures from 30° to 70℃, with maximum activity at 40℃. Furthermore, recombinant Ary423 retained more than 70% and 40% of its maximum activity after 12 h of incubation at 50℃ and 60℃, respectively, exhibiting good thermostability at high temperatures. The optimal pH for Ary423 was determined to be 8.0 and the activity of Ary423 could be slightly enhanced by Mg²⁺. The recombinant enzyme could hydrolyze sulfate ester bonds in p-nitrophenyl sulfate (NPS) and Asparagus crude polysaccharides with a specific activity of 64.8 U/mg and 25.4 U/mg, respectively. These favorable properties could make Ary423 attractive for application in the desulfating process of agar production.

• 자연과학논문집
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• 제16권1호
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• pp.1-13
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• 2005

안지오텐신(ACE) 저해제는 항고혈 효과를 갖고 있으므로 오랫동안 고혈압의 예방이나 치료에 이용되어 왔다. 본 연구는 재조합 대장균으로부터 새로운 ACE 저해제를 생산하고 정제하며 나아가 이들이 구조-기능 관P를 규명하기 위해 수행되었다. Saccharomyces cerevisiae의 ACE 저해 펩타이드 유전자를 함유하고 있는 재조합 pGEX-4T-3을 대장균 BL21(DE3)로 형질전환 시켰다. 재조합 pGEX-4T-3을 갖고 있는 대장균 BL21(DE3)로부터 생산된 Glutathione-s 전이효소(GST) 융합 단백질을 얻어서 그중 ACE저해 펩타이드를 Sephadex G-25 컬럼 크로마토그래피로 정제하였다. 정제된 ACE 저해 펩타이드는 타이로신-아스파틱엑시드-그리신-글리신-발린-페닐알라린-아르기닌-발린-타이로신-트레오닌의 서열을 가진 새로운 decapeptide이었고 ACE에 대하여 경쟁적으로 저해하였다.

• The Plant Pathology Journal
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• 제31권4호
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• pp.414-420
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• 2015

Antifungal activities of crude extractum of Nanshancha Seed Cake (NSC), to inactivate postharvest pathogens were investigated. Highest inhibitory rate was found against C. musae, C. gloeosporioides and C. papaya P.Henn, which was much stronger than that by tea saponin. Compared to tea saponin, effects of NSC extractum was relatively weak and similar on C. gloeosporioides Penzig and P. italicum. In an in vivo study, best controlling effects by NSC extractum was found with banana anthracnose disease development, which showed no inhibitory effects by tea saponin. NSC extractum controlled in vitro C. musae growth through directly inhibiting germination rate and germ tube elongation, and causing distortation, rupture and indentation of C. musae mycelium. In banana fruit subject to C. musae inoculation, higher PAL, POD, GLU and CHT activity was observed in banana fruit treated with crude NSC extractum than that of water control fruits. Current study proved the best controlling effects of crude NSC extractum in C. musae in vitro and in vivo development, which through direct inhibition of C. musae growth and increasing defense system of the banana fruit.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.660-665
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• 2003

High-level expression of Thermus caldophilus GK24 alkaline phosphatase (Tca APase) was achieved in Escherichia coli using the pET-based expression plasmids, pEAP1 and pEAP2. In the case of plasmid pEAP2, the signal peptide region of Tca APase was replaced by the PelB leader peptide of expression vector pET-22b(+). Furthermore, the expression level was somewhat higher than that of plasmid pEAPl. A rapid purification procedure of Tca APase overproduced in E. coli was developed which involved heating to denature E. coli proteins followed by HiTrap Heparin HP column chromatography. Optimal temperature and pH and $Mg^{2+}$ dependence of the recombinant Tca APase were similar to those of native enzyme isolated from T. caldophilus GK24.

• 한국약용작물학회지
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• 제21권3호
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• pp.220-228
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• 2013

This study was conducted to assess the genetic diversity and population structure of 70 amaranth accessions collected from South and Southeast Asia using 14 simple sequence repeat (SSR) markers. In total, 67 alleles were detected, with an average of 4.79 per locus. Rare alleles comprised a large portion (46.3%) of the detected alleles, and 29 unique alleles associated with rice accessions were also discovered. The mean major allele frequency (MAF), genetic diversity (GD) and polymorphic information content (PIC) of the 14 SSR loci were 0.77, 0.36, and 0.34, respectively. A model-based structural analysis revealed the presence of three subpopulations. The genetic relationships revealed by the neighbor-joining tree method were fairly consistent with the structure-based membership assignments for most of the accessions. All 70 accessions showed a clear relationship to each cluster without any admixtures. We observed a relatively low extent of genetic exchange within or among amaranth species from South and Southeast Asia. The genetic diversity results could be used to identify amaranth germplasms and so facilitate their use for crop improvement.

• Mycobiology
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• 제49권1호
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• pp.61-68
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• 2021

Agaricus bisporus, commonly known as the button mushroom, is widely cultivated throughout the world. To breed new strains with more desirable traits and improved adaptability, diverse germplasm, including wild accessions, is a valuable genetic resource. To better understand the genetic diversity available in A. bisporus and identify previously unknown diversity within accessions, a phylogenetic analysis of 360 Agaricus spp. accessions using single-nucleotide polymorphism genotyping was performed. Genetic relationships were compared using principal coordinate analysis (PCoA) among accessions with known origins and accessions with limited collection data. The accessions clustered into four groups based on the PCoA with regard to genetic relationships. A subset of 67 strains, which comprised a core collection where repetitive and uninformative accessions were not included, clustered into 7 groups following analysis. Two of the 170 accessions with limited collection data were identified as wild germplasm. The core collection allowed for the accurate analysis of A. bisporus genetic relationships, and accessions with an unknown pedigree were effectively grouped, allowing for origin identification, by PCoA analysis in this study.

• Interdisciplinary Bio Central
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• 제2권4호
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• pp.15.1-15.5
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• 2010

RIKEN BioResource Center (BRC) has collected, preserved, conducted quality control of, and distributed mouse resources since 2002 as the core facility of the National BioResource Project by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. Our mouse resources include over 5,000 strains such as humanized disease models, fluorescent reporters, and knockout mice. We have developed novel mouse strains such as tissue-specific Cre-drivers and optogenetic strains that are in high demand by the research community. We have removed all our specified pathogens from the deposited mice and used our quality control tests to examine their genetic modifications and backgrounds. RIKEN BRC is a founding member of the Federation of International Mouse Resources and the Asian Mouse Mutagenesis and Resource Association, and provides mouse resources to the one-stop International Mouse Strain Resource database. RIKEN BRC also participates in the International Gene Trap Consortium, having registered 713 gene-trap clones and their sequences in a public library, and is an advisory member of the CREATE (Coordination of resources for conditional expression of mutated mouse alleles) consortium which represents major European and international mouse database holders for the integration and dissemination of Cre-driver strains. RIKEN BRC provides training courses in the use of advanced technologies for the quality control and cryopreservation of mouse strains to promote the effective use of mouse resources worldwide.

• 한국자원식물학회지
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• 제34권4호
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• pp.311-338
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• 2021

본 연구는 경기도 김포의 최서북단 DMZ 접경지역 및 민북지역에 위치한 문수산의 산림유전자원보호구역 일대의 관속식물상과 침입외래식물 등을 파악하고 확인된 식물들의 효율적인 관리와 생물다양성 보전을 위한 기초자료로 활용하기 위하여 실시하였다. 조사는 2019년 4월부터 2020년 10월까지 총 17회에 걸쳐 실시되었다. 문수산 산림유전자원보호구역 일대의 관속식물상은 총 95과 276속 395종 13아종 33변종 3품종 444분류군이 분포하는 것으로 확인되었다. 이는 우리나라 관속식물 총 4,881분류군의 약 9.09%로 나타났다. 문수산 산림유전자원보호구역에서 출현한 한반도 특산식물은 은사시나무, 오동나무, 산이스라지, 백운산운추리 등 6종이 관찰되었다. IUCN 지정 희귀식물은 총 3분류군이 관찰되었다. 위기종(EN)은 복사앵도나무, 약관심종은 쥐방울덩굴과 창포가 출현하였다. 식물구계학적 특정식물은 초 39분류군으로 파악되었다. IV등급은 바위말발도리 등 2분류군, III등급은 고광나무, 병아리꽃나무 등 8분류군이 확인되었다. II등급은 잣나무 및 오리나무 등 총 10분류군이 관찰되었다. 조사지역에서 출현한 침입외래식물은 개망초, 달맞이꽃 및 개쑥갓 등 58분류군으로 확인되었다. 귀화율은 13.1%로 나타났으며, 도시화지수는 18.0%로 계산되었다. 본 연구에서 파악된 문수산 산림유전자원보호구역의 관속식물상과 선행연구와 비교한 결과, 최근 조사 구간이 더 넓고 다양한 경로를 조사했음에도 불구하고 선행연구에 비해 분류군 수가 감소한 것으로 파악되었다. 특히 희귀·특산식물의 수는 크게 감소하고 외래침입식물의 수는 임도 및 등산로 등으로 크게 확산한 것으로 확인되었다. 따라서 산림유전자원보호구역 내 자생식물의 보전 및 관리를 위해 산림휴식년제 등을 도입해야 할 것으로 생각된다.

• 한국산업경영시스템학회:학술대회논문집
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• 한국산업경영시스템학회 2002년도 춘계학술대회
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• pp.203-208
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• 2002

In case of the problems with multiple plants, alternative operation sequence, alternative machine, setup time, and transportation time between plants, we need a robust methodology for the integration of process planning and scheduling in supply chain. The objective of this model is to minimize the tardiness and to maximize the resource utilization. So, we propose a multi-objective model with limited-capacity constraint. To solve this model, we develope an efficient and flexible model using adaptive genetic algorithm(AGA), compared to traditional genetic algorithm(TGA)