• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.77-83
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• 1999

Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

• Fisheries and Aquatic Sciences
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• v.13 no.4
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• pp.272-277
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• 2010

Seagrasses are marine angiosperms of ecological importance in providing shelter and food to aquatic species as well as maintaining the carbon cycle on earth. Phyllospadix iwatensis is a seagrass of the family Zosteraceae and is distributed along the eastern coast of Korea. The nucleotide sequences of P. iwatensis nuclear genes encoding 18S ribosomal RNA (rRNA) and internal transcribed spacer-1 (ITS-1) were determined for molecular phylogenetic analysis. Genomic DNA was isolated from P. iwatensis and used for PCR amplification of 18S rRNA and ITS-1. Examination of the 18S rRNA sequence of P. iwatensis showed a close (99% similarity) relationship to Zostera noltii, another genus of Zosteraceae, but a distant (84% similarity) evolutionary relationship to other macroalgal Laminariales species. Further discrepancies found in ITS-1 nucleotide sequences between closely related species indicate that the sequence information could be used for species identification.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.3
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• pp.155-162
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• 2002

The cyclic amidohydrolase family enzymes, which include allantoinase, dihydroorotase, dihydropyrimidinase and (phenyl)hydantoinase, are metal-dependent hydrolases and play a crucial role in the metabolism of purine and pyrimidine in vivo. Each enzyme has been independently characterized, and thus well documented, but studies on the higher structural traits shared by members of this enzyme family are rare due to the lack of comparative study. Here, we report upon the expression in E. coli cells of maltose-binding protein (MBP)- and glutathione S-transferase (GST)-fused cyclic amidohydrolase family enzymes, facilitating also for both simple purification and high-level expression. Interestingly, the native quaternary structure of each enzyme was maintained even when fused with MBP and GST. We also found that in fusion proteins the favorable biochemical properties of family enzymes such as, their optimal pHs, specific activities and kinetic properties were conserved compared to the native enzymes. In addition, MBP-fused enzymes showed remarkable folding ability in-vitro. Our findings, therefore, suggest that a previously unrecognized trait of this family, namely the ability to functional fusion with some other protein but yet to retain innate properties, is conserved. We described here the structural and evolutionary implications of the properties in this family enzyme.

• Mycobiology
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• v.48 no.5
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• pp.331-340
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• 2020

The family Peronosporaceae, an obligate biotrophic group of Oomycota, causes downy mildew disease on many cultivated and ornamental plants such as beet, cucumber, grape, onion, rose, spinach, and sunflower. To investigate the diversity of Peronosporaceae species in Korea, we performed morphological analysis for dried plant herbariums with downy mildew infections by two largest genera, Peronospora and Plasmopara. As a result, it was confirmed that there are five species of Peronospora and two species of Plasmopara, which have been so far unrecorded in Korea, as well as rarely known in the world; Pl. angustiterminalis (ex Xanthium strumarium), Pl. siegesbeckiae (ex Siegesbeckia glabrescens), P. chenopodii-ambrosioidis (ex Chenopodium ambrosioides), P. chenopodii-ficifolii (ex Chenopodium ficifolium), P. clinopodii (ex Clinopodium cf. vulgare), P. elsholtziae (ex Elsholtzia ciliata), and P. lathyrina (ex Lathyrus japonicus). In addition, their phylogenetic relationship was inferred by molecular sequence analysis of ITS, LSU rDNA, and cox2 mtDNA. By rediscovering the seven missing species and barcoding their DNA sequences, this study provides valuable insights into the diversity and evolutionary studies of downy mildew pathogens.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.373-382
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• 2020

Candidatus Carsonella ruddii is an endosymbiont that resides in specialized cells within the body cavity of plant sap-feeding insects called psyllids. The establishment of symbiotic associations is considered one of the key factors for the evolutionary success of psyllids, as it may have helped them adapt to imbalanced food resources like plant sap. Although C. ruddii is defined as a psyllid primary symbiont, the genes for some essential amino acid pathways are absent. Complete genome sequences of several C. ruddii strains have been published. However, in-depth intra-species comparison of C. ruddii strains has not yet been done. This study therefore aimed to perform a comparative genome analysis of six C. ruddii strains, allowing the interrogation of phylogenetic group, functional category of genes, and biosynthetic pathway analysis. Accordingly, overall genome size, number of genes, and GC content of C. ruddii strains were reduced. Phylogenetic analysis based on the whole genome proteomes of 30 related bacterial strains revealed that the six C. ruddii strains form a cluster in same clade. Biosynthetic pathway analysis showed that complete sets of genes for biosynthesis of essential amino acids, except tryptophan, are absent in six C. ruddii strains. All genes for tryptophan biosynthesis are present in three C. ruddii strains (BC, BT, and YCCR). It is likely that the host may depend on a secondary symbiont to complement its deficient diet. Overall, it is therefore possible that C. ruddii is being driven to extinction and replacement by new symbionts.

• Proceedings of the Korean Information Science Society Conference
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• 2005.07b
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• pp.265-267
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• 2005

DNA 컴퓨팅은 분자 수준(molecular level)에서 연산을 수행한다. 따라서 일반적인 실리콘 기반의 컴퓨터에서와는 달리, 순차적인 연산 제어를 보장하기 어렵다는 특징이 있다. 그러나 DNA 컴퓨팅은 화학반응에 기초한 연산이기 때문에, 실험자가 의도한 연산을 많은 수의 분자에 동시에 적용할 수 있으므로 실리콘 기반의 컴퓨터와는 비교할 수 없는 병렬 연산을 구현할 수 있다. 병렬 연산을 구현하고자 할 때, 일반적으로 연산에 사용하는 모든 DNA 분자들을 대상으로 연산을 구현할 수도 있다. 그러나 전체가 아닌 일부의 분자들을 상대로 연산을 수행하는 것 역시 가능하며 이 때 자연스러운 방법으로 사용할 수 있는 방법이 배깅(Bagging)이나 부스팅(Boosting)과 같은 앙상블(ensemble) 계열의 학습 방법이다. 일반적인 부스팅과 달리 가중치를 부여하는 것이 아니라 특정 학습자(learner)를 나타내는 분자들을 증폭한다면 가중치를 분자의 양으로 표현하는 것이 가능하므로 분자 수준에서 앙상블 계열의 학습을 구현하는 것이 가능하다. 본 논문에서는 앙상블 계열의 학습 방법 중 특히 부스팅의 효과를 DNA 컴퓨팅에 응용하고자 할 때, 어떤 방법이 가능하며, 표현 과정에서 고려해야 할 사항은 어떠한 것들이 있는지 고려하고자 한다. 본 논문에서는 규모를 사전에 한정할 수 없는 진화 가능한 그래프 구조(evolutionary graph structure)를 학습할 수 있는 방법을 찾아보고자 한다. 진화 가능한 그래프 구조는 기존의 DNA 컴퓨팅 방법으로는 학습할 수 없는 문제이다. 그러나 조합 가능한 수를 사전에 정의할 수 없기 때문에 분자의 수에 상관없이 동일한 연산 시간에 문제를 해결할 수 있는 DNA 컴퓨팅의 장정을 가장 잘 발휘할 수 있는 문제이기도 하다.개별 태스크의 특성에 따른 성능 조절과 태스크의 변화에 따른 빠른 반응을 자랑으로 한다. 본 논문에선 TIB 알고리즘을 리눅스 커널에 구현하여 성능을 평가하였고 그 결과 리눅스에서 사용되는 기존 인터벌 기반의 알고리즘들에 비해 좋은 전력 절감 효과를 얻을 수 있었다.과는 한식 외식업체들이 고객들의 재구매 의도를 높이기 위해서는 한식 외식업체의 서비스요인, 식음료요인, 이벤트 요인 등을 강화함으로써 전반적인 종사원 서비스 품질과 식음료품질을 높이는 전략을 취해야 한다는 것을 시사해주고 있다. 본 연구는 대구 경북소재 한식 외식업체만을 대상으로 하여 연구를 실시하여 연구의 일반화와 한식 외식업체를 이용하는 이용 고객들이 한식 외식업체를 재방문하는 재구매 의도가 발생하는데 있어 발생하는 과정을 설명하는 종단적 연구를 실시하지 못한 한계점을 가지고 있다.아직 산업 디자인이 품질경쟁력에 크게 영향을 미치는 성숙단계에 이르지 못하였음을 의미한다. (2) 제품 디자인에게 영향을 끼치는 유의적인 변수는 연구개발력, 연구개발투자 수준, 혁신활동 수준(5S, TPM, 6Sigma 운동, QC 등)이며, 제품 디자인은 우선 품질경쟁력을 높여 간접적으로 고객만족과 고객 충성을 유발하는 것으로 추정되었다. 상기의 분석결과로부터, 본 연구는 다음과 같은 정책적 함의를 도출하였다. 첫째, 신상품 개발과 혁신을 위한 포괄적인 연구개발 프로젝트를 품질 경쟁력의 주요 결정요인(제품의 기본성능, 신뢰성, 수명(내구성) 및 제품 디자인)과 연계하여 추진해야 할 것이다. 둘째, 기업은 디자인 경영 마인드 제고와 디자인 전문인력 양성을, 대학은 디자인 현장 업무를 통하여 창의력 증진과 기획 및 마케팅 능력 교육을, 정부는 디자인 기술개발 및 디자인 교육지원의 강화를 통하여 각각 디자인 경쟁력$\righta

• Molecules and Cells
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• v.43 no.4
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• pp.331-339
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• 2020

Genetic modifications in noncoding regulatory regions are likely critical to human evolution. Human-accelerated noncoding elements are highly conserved noncoding regions among vertebrates but have large differences across humans, which implies human-specific regulatory potential. In this study, we found that human-accelerated noncoding elements were frequently coupled with DNase I hypersensitive sites (DHSs), together with monomethylated and trimethylated histone H3 lysine 4, which are active regulatory markers. This coupling was particularly pronounced in fetal brains relative to adult brains, non-brain fetal tissues, and embryonic stem cells. However, fetal brain DHSs were also specifically enriched in deeply conserved sequences, implying coexistence of universal maintenance and human-specific fitness in human brain development. We assessed whether this coexisting pattern was a general one by quantitatively measuring evolutionary rates of DHSs. As a result, fetal brain DHSs showed a mixed but distinct signature of regional conservation and outlier point acceleration as compared to other DHSs. This finding suggests that brain developmental sequences are selectively constrained in general, whereas specific nucleotides are under positive selection or constraint relaxation simultaneously. Hence, we hypothesize that human- or primate-specific changes to universally conserved regulatory codes of brain development may drive the accelerated, and most likely adaptive, evolution of the regulatory network of the human brain.

• Journal of Microbiology
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• v.44 no.6
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• pp.655-659
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• 2006

Phylogenetic studies of nef, pol, and env gene sequences of HIV-1 isolated from Koreans suggested the presence of a Korean clade in which Korean sequences are clustered to the exclusion of foreign sequences. We attempted to identify and characterize the Korean clade using all vif gene sequences isolated from Koreans registered in the NCBI GenBank database (n = 233). Most (77 %) of the Korean isolates belonged to the Korean clade as a large subcluster in subtype B, designated the Korean clade subtype B ($K_{C}B$). $K_{C}B$ sequences were relatively homogenous compared to Korean subtype B sequences that did not belong to the $K_{C}B$ (non-Korean clade subtype B; $NK_{C}B$). Comparison of amino acid frequencies of $K_{C}B$ and $NK_{C}B$ sequences revealed several positions where the amino acid frequencies were significantly different. These amino acid residues were critical in separating $K_{C}B$ from $NK_{C}B$ or from foreign sequences, since substitution of these amino acids in $K_{C}B$ with the $NK_{C}B$ amino acids relocated the $K_{C}B$ sequences to $NK_{C}B$, and vice versa. Further analyses of $K_{C}B$ will help us to understand the origin and evolutionary history of $K_{C}B$.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1680-1685
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• 2015

Since ancient days, domestic horses have been closely associated with human civilization. Today, horse racing is an important industry. Various genes involved in energy production and muscle contraction are differentially regulated during a race. Among them, creatine kinase (CK) is well known for its regulation of energy preservation in animal cells. CK is an iso-enzyme, encoded by different genes and expressed in skeletal muscle, heart, brain and leucocytes. We confirmed that the expression of CK-M significantly increased in the blood after a 30 minute exercise period, while no considerable change was observed in skeletal muscle. Analysis of various tissues showed an ubiquitous expression of the CK-M gene in the horse; CK-M mRNA expression was predominant in the skeletal muscle and the cardiac muscle compared to other tissues. An evolutionary study by synonymous and non-synonymous single nucleotide polymorphism ratio of CK-M gene revealed a positive selection that was conserved in the horse. More studies are warranted in order to develop the expression of CK-M gene as a biomarker in blood of thoroughbred horses.

• Development and Reproduction
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• v.10 no.2
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• pp.127-133
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• 2006

[ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.