• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2005년도 춘계 학술발표대회
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• pp.143-143
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• 2005

• 한국해양바이오학회지
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• 제2권3호
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• pp.168-173
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• 2007

of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics. To analyze the transcriptome of olive flounder, Paralichthys olivaceus, we have conducted EST analysis using cDNA libraries made from muscle of P. olivaceus. Redundant ESTs were assembled into overlapping contigs by using the assembly program ICAtools software. We found that the 221 ESTs were composed of 21 clusters and 35 singletons, suggesting that the overall redundancy of the library was 74.7%. Of the 221 clones, 218 clones (98.6%) were identified as known genes by BLAST searches and 3 clones (1.4%) did not match to any previously described genes. Based on major functions of their encoded proteins, the identified clones were classified into 13 broad categories. Sequence analysis of the ESTs revealed the presence of microsatellite-containing genes which may be valuable for further gene mapping studies. This study contributes to the identification of many EST clones that could be useful for genetics and developmental biology of olive flounder.

• BMB Reports
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• 제44권4호
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• pp.238-243
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• 2011

We generated 16,993 expressed sequence tags (ESTs) from two libraries containing full-length cDNAs from the brain and liver of the Korean Jindo dog. An additional 365,909 ESTs from other dog breeds were identified from the NCBI dbEST database, and all ESTs were clustered into 28,514 consensus sequences using StackPack. We selected the 7,305 consensus sequences that could be assembled from at least five ESTs and estimated that 12,533 high-quality single nucleotide polymorphisms (SNPs) were present in 97,835 putative SNPs from the 7,305 consensus sequences. We identified 58 Jindo dog-specific SNPs in comparison to other breeds and predicted seven synonymous SNPs and ten non-synonymous SNPs. Using PolyPhen, a program that predicts changes in protein structure and potential effects on protein function caused by amino acid substitutions, three of the non-synonymous SNPs were predicted to result in changes in protein function for proteins expressed by three different genes (TUSC3, ITIH2, and NAT2).

• Journal of Plant Biotechnology
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• 제1권1호
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• pp.46-55
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• 1999

The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.

• 한국잠사곤충학회지
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• 제38권1호
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• pp.13-18
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• 1996

곤충의 다양한 기능해석을 유전자 수준에서 수행하기 위하여 주요 익충인 누에를 대상으로 유전 자원 확보를 시도하였다. 우선 5령의 누에유충에서 cDNA 유전자 은행을 제작하여 1.3 X 106개의 cDNA 유전자원을 확보하였다. 누에유충의 cDNA 유전자 은행에서 무작위로 plaques을 선정하였고, 이를 플라스미드로 전환하여 SK primer를 이용한 부분 염기서열을 결정하였다. 결정된 cDNA 클론의 부분 염기서열을 GenBank 데이타베이스에서 검색하여 37개의 발현 유전자 꼬리표를 생산하였다. 이들 중 15개는 데이터베이스와의 비교부위가 150bp 이상이고 DNA 상동성이 약 60% 이상으로 비교적 높은 DNA 상동 유의성을 나타내는 것으로 혈림프에서 발견되는 수종의 저장 단백질, 곤충의 기동성, 체벽 형성, 효소 및 초파리의 돌연변이 유전자형과 유사한 종류들이었다. 또한 15개의 발현 유전자 꼬리표 중 누에에서 밝혀진 것은 3종이고 그 나머지는 누에에서 처음 밝혀진 것으로 이 클론에 대한 정확한 동정이 요구된다.

• 한국양식학회지
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• 제20권4호
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• pp.248-254
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• 2007

멸종위기 천연기념물 어류 어름치(Hemibarbus mylodon)를 대상으로한 어름치 유전자 은행 구축 연구의 일환으로 뇌, 소화관, 근육, 간, 신장, 난소 및 정소 조직으로부터 expressed sequence tag (EST) library들을 구축하고 발현 유전자의 탐색을 실시하였다. EST 탐색을 통해 총 3,383개의 발현 유전자 염기서열 단편을 확보하였고 이들로부터 1,354개의 EST를 포함하는 총 333개의 contig들이 형성됨으로써 비교적 높은 빈도(69.8%)의 unigene 확보율을 나타내었다. EST의 조직 별 출현 양상은 orthologue들과의 상동성 정도 및 유추 기능의 대분류를 기준으로 분석할 때 각 조직들은 서로 다른 특징을 나타내었다. 어름치에서 발굴된 EST들은 zebrafish의 유전자들과 가장 높은 match 빈도를 나타내었다. 본 연구를 통해 확보된 EST library들과 염기서열 정보는 본 종의 장외 복원을 위한 효율적인 인공증식 기술 개발에 유용한 기초 정보로 활용될 수 있을 것이다.

• Fisheries and Aquatic Sciences
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• 제10권3호
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• pp.119-126
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• 2007

The analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and the development of resources for functional genomics. To analyze the transcriptome of the abalone Haliotis discus hannai, we conducted EST analysis using seven cDNA libraries made from gill, gut, hepatopancreas, skin, muscle, testis, and ovary. Redundant ESTs were assembled into overlapping contiguous sequences using the assembly program ICAtools. We found that the total 1,393 ESTs formed 135 clusters and 951 singletons, indicating that the overall redundancy of the library was 22%. Of the 1,393 clones, BLAST identified 1,278 clones (91.7%) as known genes; 115 clones (8.3%) did not match any previously described gene. Based on the major functions of their encoded proteins, the identified clones were classified into 16 broad categories. Sequence analysis revealed the presence of micro satellite-containing genes that may be valuable for further gene mapping studies. This study contributes to the identification of numerous EST clones that can be applied to further clarifying the genetics and developmental biology of abalone.

• Journal of Plant Biotechnology
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• 제37권4호
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• pp.472-482
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• 2010

For the massive screening of the genes related to oxidative stress, a cDNA library was constructed from hot pepper (Capsicum annuum L. cv. Nockkwang) leaves treated with methyl viologen. From this library, 1,589 cDNA clones were sequenced from their 5' ends. The sequences were clustered into 1,252 unigenes comprised of 152 contigs and 1,100 singletons. Similarity search against NCBI protein database identified 1,005 ESTs (80.3%) as Known, 197 ESTs (15.7%) as Unknown, and 50 ESTs (3.99%) as No hit. In the ESTs, oxidative stress-related genes such as ascorbate peroxidase, catalase, and osmotin precursor were highly expressed. The cDNA microarray containing 1,252 unigenes was constructed and used to analyze their expression upon methyl viologen treatment. Analyses of the hybridization revealed that various stress-related genes such as peroxidase, tyrosine aminotransferase, and omega-6 fatty acid desaturase, were induced and some metabolism related genes such as aldolase and ketol-acid reductoisomerase, were repressed by methyl viologen treatment, respectively. The information from this study will be used for further study on the functional roles of oxidative stress-related genes and signaling network of oxidative stress in hot pepper.

• The Plant Pathology Journal
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• 제30권4호
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• pp.425-431
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• 2014

Leaf scald caused by the infection of Rhynchosporium secalis, is a worldwide crop disease resulting in significant loss of barley yield. In this study, a systematic sequencing of expressed sequence tags (ESTs) was chosen to obtain a global picture of the assembly of genes involved in pathogenesis. To identify a large number of plant ESTs, which are induced at different time points, an amplified fragment length polymorphism (AFLP) display of complementary DNA (cDNA) was utilized. Transcriptional changes of 140 ESTs were observed, of which 19 have no previously described function. Functional annotation of the transcripts revealed a variety of infection-induced host genes encoding classical pathogenesis-related (PR) or genes that play a role in the signal transduction pathway. The expression analyses by a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) revealed that Rar1 and Rpg4 are defense inducible genes, and were consistent with the cDNA-AFLP data in their expression patterns. Hence, the here presented transcriptomic approach provides novel global catalogue of genes not currently represented in the EST databases.

• Parasites, Hosts and Diseases
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• 제39권2호
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• pp.151-160
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• 2001

Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface Proteins , enzymes for DNA, energy Production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba.