• Title/Summary/Keyword: Activity Sequencing

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Production of DNA polymerase from Thermus aquaticus in recombinant Escherichia coli

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.245-249
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    • 2014
  • Among dozens of DNA polymerases cloned from thermophilic bacteria, Taq DNA polymerase from Thermus aquaticus has been most frequently used in polymerase chain reaction (PCR) that is being applied to gene cloning, DNA sequencing, gene expression analysis, and detection of infectious and genetic diseases. Since native Taq DNA polymerase is expressed at low level in T. aquaticus, recombinant Escherichia coli system was used to produce Taq DNA polymerase in a large amount. Taq DNA polymerase was expressed as a soluble form under the control of tac promoter in E. coli, and purified by heat treatment and ion exchange chromatographies. The purified Taq DNA polymerase was nearly homogeneous and exhibited a similar DNA amplification activity with a commercial Taq DNA polymerase.

Draft genome sequence of Lactobacillus reuteri KLR3004 from a fattening pig (비육돈 분변으로부터 분리한 Lactobacillus reuteri KLR3004 유산균주의 유전체 분석)

  • Park, Jongbin;Lee, Jun-Yeong;Jin, Gwi-Deuk;Kim, Eun Bae
    • Korean Journal of Microbiology
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    • v.53 no.2
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    • pp.146-148
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    • 2017
  • We sequenced the genome of Lactobacillus reuteri KLR3004 strain isolated from a fattening pig in South Korea. The sequences were assembled into a draft genome containing 1,996,237 bp with a G+C content of 38.75% and 1,837 predicted protein-coding sequences in 149 contigs.

A FRAMEWORK FOR SIMULATING CONSTRUCTION PROCESSES FOR OPTIMIZING THE FLOOR CONSTRUCTION CYCLE USING BIM

  • Seung-Jun Ahn;Hyun-Soo Lee;Moonseo Park
    • International conference on construction engineering and project management
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    • 2009.05a
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    • pp.835-840
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    • 2009
  • Lately, Building Information Modeling (BIM) emerges as the most promising technology, now is expected to bring a great deal of improvement of productivity in every aspect of the construction industry. One of the BIM based scheduling is to use BIM model as a base for applying to schedule analysis and simulation tools. This type of tools may incorporate a various types of information such as the building model, construction method information, resource information, productivity information, rules and constraints to optimize activity sequencing. This paper proposes a framework of BIM based simulating system which can be used to optimize construction processes, especially for the floor construction cycle. For the purpose, all of the necessary components of the system will be defined and represented, and next an algorithm will be introduced to demonstrate the principle of simulating operation. The benefits of this technique are basically two : to test and optimize construction methods in respect of the construction duration and to reduce the floor construction cycle.

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Characterization of Root Transcriptome among Korean Ginseng Cultivars and American Ginseng using Next Generation Sequencing (차세대염기서열 분석을 이용한 고려인삼과 미국삼의 전사체 분석)

  • Jo, Ick Hyun;Kim, Young Chang;Lee, Seung Ho;Kim, Jang Uk;Kim, Sun Tae;Hyun, Dong Yun;Kim, Dong Hwi;Kim, Kee Hong;Kim, Hong Sig;Chung, Jong Wook;Bang, Kyong Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.5
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    • pp.339-348
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    • 2014
  • The transcriptomes of four ginseng accessions such as Cheonryang (Korean ginseng cultivar), Yunpoong (Korean ginseng cultivar), G03080 (breeding line of Korean ginseng), and P. quinquefolius (American ginseng) was characterized. As a result of sequencing, total lengths of the reads in each sample were 156.42 Mb (Cheonryang cultivar), 161.95 Mb (Yunpoong cultivar), 165.07 Mb (G03080 breeding line), and 166.48 Mb (P. quinquefolius). Using a BLAST search against the Phytozome databases with an arbitrary expectation value of 1E-10, over 20,000 unigenes were functionally annotated and classified using DAVID software, and were found in response to external stress in the G03080 breeding line, as well as in the Cheonryang cultivar, which was associated with the ion binding term. Finally, unigenes related to transmembrane transporter activity were observed in Cheonryang and P. quinquefolius, which involves controlling osmotic pressure and turgor pressure within the cell. The expression patterns were analyzed to identify dehydrin family genes that were abundantly detected in the Cheonryang cultivar and the G03080 breeding line. In addition, the Yunpoong cultivar and P. quinquefolius accession had higher expression of heat shock proteins expressed in Ricinus communis. These results will be a valuable resource for understanding the structure and function of the ginseng transcriptomes.

Isolation of Simiduia sp. SH-2 and Characterization of Its β-Agarase (한천분해세균 Simiduia sp. SH-2 균주의 분리 및 β-agarase의 특성조사)

  • Lee, Dong-Geun;Kim, Geun-Dae;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.32 no.10
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    • pp.778-783
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    • 2022
  • This study isolated a new agarase-producing bacterium and characterized its agarase. A new agar-degrading strain was isolated from the seashore of Namhae in Gyeongnam province, Korea, and was purely cultured using the Marine Agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-2 after 16S rRNA gene sequencing. The crude agarase was obtained from the culture medium of the Simiduia sp. SH-2 strain, and the agar-degrading activity was measured. The highest level of activity of the Simiduia sp. SH-2-derived agar-degrading enzyme was 625 U/l. Agar degradation activity was most significant at 40℃ and pH 7.0. Compared to the activity at 40℃, the relative activity was 31% at 20℃ and 71% at 30℃. Compared to the activity at pH 7.0, the relative activity was 94% and 89% at pH 6.0 and pH 8.0, respectively. Residual activity was greater than 96% after exposure to 20℃ and 30℃ for 2 hr and more than 49% after exposure to 40℃ for 2 hr. Simiduia sp. SH-2 was identified as a strain producing β-agarase that creates neoagarooligosaccharides, such as neoagarotetraose and neoagarohexaose. Therefore, the Simiduia sp. SH-2 strain and its β-agarase are expected to be useful functional material producers in the food, cosmetic, and pharmaceutical industries.

Functional analysis of SH3 domain containing ring finger 2 during the myogenic differentiation of quail myoblast cells

  • Kim, Si Won;Lee, Jeong Hyo;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1183-1189
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    • 2017
  • Objective: Owing to the public availability of complete genome sequences, including avian species, massive bioinformatics analyses may be conducted for computational gene prediction and the identification of gene regulatory networks through various informatics tools. However, to evaluate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite. Methods: Due to a lack of quail genomic sequence information, we first identified the partial genomic structure and sequences of the quail SH3 domain containing ring finger 2 (SH3RF2) gene. Subsequently, SH3RF2 was knocked out using clustered regularly interspaced short palindromic repeat/Cas9 technology and single cell-derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation. Results: Through a T7 endonuclease I assay and genotyping analysis, we established an SH3RF2 knockout (KO) QM7#4 subline with 61 and 155 nucleotide deletion mutations in SH3RF2. After the induction of myotube differentiation, the expression profiles were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis. Conclusion: We did not detect any statistically significant role of SH3RF2 during myotube differentiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.

Isolation and Identification of Lactic Bacteria Containing Superior Activity of the Bile Salts Deconjugation (담즙산 분해능이 뛰어난 젖산균의 분리 및 동정)

  • 하철규;조진국;채영규;허강칠
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.164-170
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    • 2004
  • The purpose of this study is to isolate probiotic lactic acid bacteria (LAB) that produced bile salts hydrolase. One hundred twenty strains were initially isolated from human feces. Based on their resistance of acid, tolerances of bile salts, and inhibitory activity against Escherichia coli, five strains were selected. A strain producing highest activity of bile salts hydrolase was identified as Lactoacillus plantarum using API carbohydrate fermentation pattern and 16S rRNA sequences, and named CK102. Lactobacillus plantarum CK102 survived at a level of 1.36${\times}$10$\^$8/ CFU/$m\ell$ in pH 2 buffer for 6 h and showed exhibited excellent bile tolerance. When L plantarum CK102 was cultured with E. coli in MRS broth, no viable cells of E. coli was detected after 18 h fermentation. These results suggest that Lactobacillus plantarum CK 102 may be commercially used for the probiotic culture.

Identification of Endophytic Bacteria in Panax ginseng Seeds and Their Potential for Plant Growth Promotion (인삼종자로부터 분리된 내생균의 동정과 식물생장 촉진 관련 활성의 평가)

  • Um, Yurry;Kim, Bo Ra;Jeong, Jin Ju;Chung, Chan Moon;Lee, Yi
    • Korean Journal of Medicinal Crop Science
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    • v.22 no.4
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    • pp.306-312
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    • 2014
  • Endophytes are microorganisms that live in the internal tissues of plants without harming the host plants. In this symbiotic relationship, the host plants provide nutrients and shelter to the endophytes, in turn, endophytes can promote the growth of host plants and act as a biological control agents against plant pathogens. Plant-microbe interactions like this are noted for natural methods for sustainable agriculture and environmental conservation. However, in spite of the infinite potential, there are only a few reports on the endophytes present in ginseng. In this study, we isolated and identified the endophytes from Panax ginseng seeds and evaluated the biological activities (IAA production ability, nitrogen fixation ability, phosphate solubilization capacity, siderophore production ability, and antifungal activities) of the endophyte isolates. Eight different endophytes were identified by 16S rRNA sequencing. Most of the endophytes have antibiotic and plant growth promoting (PGP) activities. Particularly, PgSEB5-37E have the highest antibiotic activity, both PgSEB5-37B and PgSEB5-37H have high PGP traits such as an abilities to produce IAA, solubilize phosphate and fix nitrogen. These results indicated that the endophytes from P. ginseng seeds may have applicable value to many industries. In order to use the isolated endophytes, quantitative analysis and field tests are needed to be performed.

Study of Distance Relationships among Domestic Radish (Raphanus sativus L.) by Analyzing its Anti-fungal Protein Gene. (항 곰팡이 단백질 유전자 분석에 의한 국내 무 품종간 유연성에 관한 연구)

  • Hwang, Cher-Won
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1294-1297
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    • 2007
  • To define diversity of domestic radish, we analysis genetic relationship of anti-fungal protein genes from several domestic radish (Raphanus sativus L.) seeds. We have isolated from domestic radish (Baekwoon) anti-fungal protein named RAP[12]. In this report, we isolate RNAs and raw protein from radish seeds then, RT-PCR analysis was done with another known anti-fungal sequences of radish from Gene Bank/EMBL and anti-fun- gal, anti-yeast activity were done against Bot교tis cenerea, Saccharomyces cerevisiaeι Candida albicans with it's raw proteins. The anti-fungal activity was shown used all seeds but anti-yeast activity was shown only two seeds (Myungsan, Baekwoon). RT-PCR products (about 0.2 Kb) were not shown only two seeds. To identify the sequencing relationship of the domestic radish, we have cloned and sequenced RAP genes of the radish and analysis the sequence relationship with clustalw program. Thus we report the result that there are some different relationship between domestic radish and known other radish's anti- fungal protein[15].

Properties of a Fibrinolytic Enzyme Secreted by Bacillus amyloliquefaciens RSB34, Isolated from Doenjang

  • Yao, Zhuang;Liu, Xiaoming;Shim, Jae Min;Lee, Kang Wook;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.9-18
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    • 2017
  • Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity ($83.23mU/{\mu}l$) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The $K_m$ and $V_{max}$ values of recombinant AprE34 were $0.131{\pm}0.026mM$ and $16.551{\pm}0.316{\mu}M/l/min$, respectively, when measured using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.