• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.254-254
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• 2022

Astragalus membranaceus (A. membranaceus) has traditionally been used as a medicinal plant in East Asia for the treatment ofvarious diseases. A. membranaceus belongs to the legume family and is known to be rich in substances such as flavonoids and saponins. Recent pharmacological studies of A. membranaceus have shown that the plant has immunomodulatory, anti-oxidant, anti-cancer, and anti-inflammatory effects. However, knowledge of major biosynthetic pathways in A. membranaceu is still lacking. Recently developed sequencing techniques enable high-quality transcriptome analysis in plants, which is recognized as an important part in elucidating the regulatory mechanisms of many plant secondary metabolic pathways. However, it is difficult to predict the number of transcripts because plant transcripts contain a large number of isoforms due to alternative splicing events, which can vary depending on the assembly platform used. In this study, we constructed three unigene sets using Pac-Bio isoform sequencing, Trinity and rnaSPAdes assembly for detailed transcriptome analysis mA. membranaceus. Furthermore, all genes involved in the flavonoid biosynthetic pathway were searched from three unigene sets, and structural comparisons and expression profiles between these genes were analyzed. The isoflavone synthesis was active in most tissues. Flavonol synthesis was mainly active in leaves and flowers, and anthocyanin synthesis was specific in flowers. Gene structural analysis revealed structural differences in the flavonoid-related genes derived from the three unigene sets. This study suggests the need for the application of multiple unigene sets for the analysis of key biosynthetic pathways in plants.

• Journal of Microbiology and Biotechnology
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• 제19권8호
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• pp.774-780
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• 2009

Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. To create a genetic database and isolate genes regulated during mushroom development, cDNA libraries were constructed from three developmental stages of mycelium, primordium, and fruit body in F. velutipes. We generated a total of 5,431 expressed sequence tags (ESTs) from randomly selected clones from the three cDNA libraries. Of these, 3,332 different unique genes (unigenes) were consistent with 2,442 (73%) singlets and 890 (27%) contigs. This corresponds to a redundancy of 39%. Using a homology search in the gene ontology database, the EST unigenes were classified into the three categories of molecular function (28%), biological process (29%), and cellular component (6%). Comparative analysis found great variations in the unigene expression pattern among the three different unigene sets generated from the cDNA libraries of mycelium, primordium, and fruit body. The 19-34% of total unigenes were unique to each unigene set and only 3% were shared among all three unigene sets. The unique and common representation in F. velutipes unigenes from the three different cDNA libraries suggests great differential gene expression profiles during the different developmental stages of F. velutipes mushroom.

• 원예과학기술지
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• 제29권2호
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• pp.116-122
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• 2011

본 연구에서는 지리산에서 자생하는 한국 특산종인 노각나무(Stewartia koreana Nakai)의 EST library를 제작하고 서열을 분석하였다. 노각나무의 유엽을 재료로 cDNA library 만들었고 1,392개의 cDNA에 대한 부분 서열 분석을 진행하였다. EST와 unigene 서열의 분석은 컴퓨터를 기반으로한 filtering과 수작업 그리고 NCBI의 BLAST 분석을 통해 수행하였다. 벡터 서열과 100bp 이하의 서열을 제거한 후 1,301개의 EST를 분석하였다. 전체 150개의 contig와 743개의 singleton을 분리하여 총 893개의 unigene을 분리해냈으며 서열 분석을 통해 95개의 microsatellite를 확인하였다. NCBI 데이터베이스의 BLASTX로 상동성을 검색한 결과 EST의 65%는 기능을 알고 있는 유전자와 11.6%의 EST는 아직까지 기능이 보고되지 않은 유전자와 높은 상동성을 보였다. 남아 있는 23.2%의 EST는 기존에 데이터베이스에 보고된 유전자와 상동성을 보이지 않는 유전자로 밝혀졌다. 다양한 데이터베이스를 기반으로 한 유사성 기반 기능 분석은 노각나무의 EST가 포도나무와 포플러와 높은 유사성을 보인 것을 확인하였다. 기능에 따른 분류에 있어 molecular function은 nucleotide binding, biological process는 transport, cellular component는 plastid가 가장 높은 비율로 나왔다. 본 연구를 통해 얻어진 EST 자료는 노각나무의 새로운 유전자원에 대한 연구의 기본 자료로 유용하게 활용될 것이다.

• BMB Reports
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• 제41권12호
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• pp.875-880
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• 2008

Mbu-3 is a novel mouse brain unigene that was identified by digital differential display. In this study, expression of the gene was chased through developmental stages and the protein product was identified in the brain. The cDNA sequence was 3,995-bp long and contained an ORF of 745 AA. Database searches revealed that the chicken SST273 gene containing LRR- and Ig-domain was an mbu-3 orthologue. Tissue specificity for the gene was examined in embryos and in brains at post-natal and adult stages. During the embryonic stages, mbu-3 was localized to the central nervous system in the brain and spinal cord. In the early post-natal stages, the gene was evenly expressed in the brain. However, with aging, expression was confined to specific regions, particularly the hippocampus. The protein was approximately 95 kDa as determined by Western blot analysis of brain extracts.

• 농업과학연구
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• 제38권1호
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• pp.11-16
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• 2011

In this study, we report the generation and analysis of a total of 1,211 expressed sequence tags (ESTs) from Pinus koraiensis. A cDNA library was generated from the young leaf tissue and a total of 1,211 cDNA were partially sequenced. EST and unigene sequence quality were determined by computational filtering, manual review, and BLAST analyses. In all, 857 ESTs were acquired after the removal of the vector sequence and filtering over a minimum length 50 nucleotides. A total of 411 unigene, consisting of 89 contigs and 322 singletons, was identified after assembling. Also, we identified 77 new microsatellite-containing sequences from the unigenes and classified the structure according to their repeat unit. According to homology search with BLASTX against the NCBI database, 63.1% of ESTs were homologous with known function and 22.2% of ESTs were matched with putative or unknown function. The remaining 14.6% of ESTs showed no significant similarity to any protein sequences found in the public database. Gene ontology (GO) classification showed that the most abundant GO terms were transport, nucleotide binding, plastid, in terms biological process, molecular function and cellular component, respectively. The sequence data will be used to characterize potential roles of new genes in Pinus and provided for the useful tools as a genetic resource.

• 원예과학기술지
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• 제28권6호
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• pp.1039-1050
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• 2010

당근은 서양뿐만 아니라 중국 및 한국과 같은 아시아 전역에서 요리로 많이 이용되는 유용한 작물 중 하나이다. 그러나 당근 종자 표면에는 모(毛)가 존재하고 이 종모는 발아율을 증가시키기 위해 제거해야 한다. 더욱이 종모 처리는 시간과 인력 및 자본의 소비와 같은 추가적인 손실을 동반하였다. 이러한 문제점을 방지하기 위해 단모종자를 이용하여 모형성과 관련된 유전자의 연구가 필요하다. 당근 종모의 발달은 2차 세포벽의 합성단계 동안 cellulose의 합성 과정과 연관되어 있음을 바탕으로, EST profiling을 통해 종모와 관련된 유전자를 탐색하고자 하였다. 당근 종모 형성에 관련된 유전자 발현을 조사하기 위해, 성숙 초기 단계의 단모종자 659-1개체와 유모종자 677-14개체를 이용하여 cDNA library를 구축하였다. 단모종자 659-1개체와 유모종자 677-14개체에서 확보된 EST 염기서열의 NCBI database BLASTX 분석을 통한 EST profiling 결과, 172개와 224개의 unigene은 이미 알려진 단백질 염기서열과 상동성을 보였으며 나머지 233개와 192개의 unigene은 확인되지 않는 유전자들이었다. EST는 추정되는 기능에 따라 16개의 category로 그룹화되었다. 전체 EST 중 29개의 unigene이 2차 세포벽 합성 단계 동안 cellulose의 합성 pathway상의 종모 형성을 조절하는 유전자로 추정되며, 실제로 종모 발달과 관련된 14개의 unigene이 유모종자 계통에서만 발견되었다.

• 한국작물학회지
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• 제55권2호
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• pp.151-158
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• 2010

지금까지 양구슬냉이의 유전정보는 거의 연구되지 않았으므로 우리는 양구슬냉이의 잎으로부터 cDNA library를 제작하고 발현유전자의 종류와 기능별 분류를 조사하였다. 그 결과를 요약하면 다음과 같다. 1. cDNA library에서 1334개 의 클론들을 얻었고 삽입된 단편들의 염기서열의 평균길이는 736bp였다. 우리는 1269개의 high-quality expressed sequence tags (ESTs) 서열을 얻었다. 이러한 EST의 클러스터 분석결과 고유 염기서열(unigene)을 가진 유전자의 수는 851개를 나타냈다. 2. Unigene 476개는 GeneBank에 기능이 알려진 유전자들과 고도의 상동성을 나타내었다. 다른 375개의 unigene들은 기능이 알려지지 않은 것들이었다. 나머지 63개는 NCBI데이터베이스에 어떤 유전자와도 상동성을 보이지 않았고 이러한 유전자들은 아마도 양구슬냉이의 잎에서 발현되는 새로운 유전자일 것으로 보인다. 3. 데이터베이스에서 상동성을 나타낸 EST들을 기능별 주석에 따라서 17개의 카테고리로 분류하였다. 대표적으로 가장 분포도가 높은 카테고리는 결합 기능 또는 보조인자 요구의 단백질(27%), 대사(11%), 세포 소기관 위치(11%), 세포수송과 수송기관 그리고 수송 경로(7%), 에너지(6%), 대사와 단백질 기능의 조절(6%) 등이 있다. 이러한 우리의 연구 결과는 양구슬냉이의 유용한 유전적 자원과 전반적인 mRNA 발현 정보를 제공해 줌으로써 대체 에너지 작물로 떠오르는 양구슬냉이의 다양한 분자적 연구에 기여할 것으로 사료된다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.253-253
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• 2022

Codonopsis lanceolata (C. lanceolata) has been widely used in East Asia as a traditional medicine to treat various diseases such as bronchitis, convulsions, cough, obesity, and hepatitis. C. lanceolata belonging to Campanulaceae contains bioactive compounds such as polyphenols, saponins, and steroids. However, despite the pharmacological significance of C. lanceolata, the genetic information of this plant is limited and there are few studies of its transcriptome. In this study, we constructed a unigene set of C. lanceolata using Pac-Bio sequencing. Furthermore, the reads generated from Pac-bio and Illumina sequencing were mixed and assembled using rnaSPAdes. All genes involved in the triterpenoid pathway, a major bioactive compounds of C. lanceolata, were searched from the two unigene sets and the expression profiles of these genes were analyzed. The results showed that lupeol, beta-amyrin, and dammarenediol synthesis genes were activated in the leaves and roots of C. lanceolata. In particular, the expression of genes related to lupeol synthesis was relatively high, suggesting that the main triterpenoid of C. lanceolata is lupeol. Transcriptome studies related to lupeol synthesis in C. lanceolata have been rarely reported. Lupeol has been reported to have pharmacological effects such as anti-inflammatory, anti-cancer, and anti-bacterial. This study suggests the importance of C. lanceolata as a lupeol producing plant.

• Journal of Microbiology
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• 제45권6호
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• pp.541-546
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• 2007

Red algae are distributed globally, and the group contains several commercially important species. Griffithsia okiensis is one of the most extensively studied red algal species. In this study, we conducted expressed sequence tag (ESTs) analysis and synonymous codon usage analysis using cultured G. okiensis samples. A total of 1,104 cDNA clones were sequenced using a cDNA library made from samples collected from Dolsan Island, on the southern coast of Korea. The clustering analysis of these sequences allowed for the identification of 1,048 unigene clusters consisting of 36 consensus and 1,012 singleton sequences. BLASTX searches generated 532 significant hits (E-value <$10^{-4}$) and via further Gene Ontology analysis, we constructed a functional classification of 434 unigenes. Our codon usage analysis showed that unigene clusters with more than three ESTs had higher GC contents (76.5%) at the third position of the codons than the singletons. Also, the majority of the optimal codons of G. okiensis and Chondrus crispus belonging to Bangiophycidae were G-ending, whereas those of Porphyra yezoensis belonging to Florideophycidae were G-ending. An orthologous gene search for the P. yezoensis EST database resulted in the identification of 39 unigenes commonly expressed in two rhodophytes, which have putative functions for structural proteins, protein degradation, signal transduction, stress response, and physiological processes. Although experiments have been conducted on a limited scale, this study provides a material basis for the development of microarrays useful for gene expression studies, as well as useful information for the comparative genomic analysis of red algae.

• 농업과학연구
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• 제43권1호
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• pp.33-39
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• 2016

Brassicaceae consists of important species that have significant amounts of metabolites, and many studies have been carried out in order to understand the mechanism that improves the content of these metabolites. In Brassicacea, Cardamine manshurica Nakai is one of the important edible plants and is rich in oil, fiber, and various nutrients. In this study, we constructed cDNA library using leaves from 4 week-old plants and analyzed the ESTs of C. manshurica Nakai. One thousand thirty-nine ESTs were discovered which assembled to form 468 unigenes. The latter contained 116 contigs and 352 singletons. Similarity search of these ESTs with BLASTX revealed similarities with Arabidopsis thaliana 285 (31.9%), Arabidopsis lyrata 172 (19.3%), Capsella rubella 162 (18.1%), and Eutrema salsugineum 137 (15.3%). ESTs were functionally categorized into molecular function, biological process, and cellular component, and each category took 10.6%, 58.5%, and 30.9%, respectively. The functional analysis also found that 94.9% of ESTs showed at least one GO ID. Microsatellite analysis of 468 unigene sequences revealed 225 structures of which Di-, Tri-, Tetra-, Penta-repeats were 35.6% (80/225), 63.1% (142/225), 0.9% (2/225), and 0.4% (1/225), respectively. The results from our study can be a valuable resource for Cardamine research.