• Korean Journal of Ichthyology
• /
• v.18 no.4
• /
• pp.283-292
• /
• 2006

Expressed sequence tag (EST) analysis was conducted using a complementary DNA (cDNA) library made from the kidney mRNA of olive flounder (Paralichthys olivaceus). In the survey of 390 ESTs chosen from the kidney cDNA library, 250 ESTs showed significant homology to previously described genes while 140 ESTs were unidentified or novel. Comparative analysis of the 250 identified ESTs showed that 14 (5.6%) clones were representing 11 unique genes identified as homologous to the previously reported olive flounder ESTs, 198 (79.2%) clones representing 160 unique genes were identified as orthologs of known genes from other organisms, and orthologs were established for 38 (15.2%) clones representing 37 genes of known sequences with unknown functions. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immunerelated genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

• Journal of fish pathology
• /
• v.23 no.3
• /
• pp.429-440
• /
• 2010

We constructed a rock bream (Oplegnathus fasciatus) gill cDNA library and a total of 1450 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 1450 EST clones, 1022 EST clones showed significant homology to previously described genes while 428 ESTs were unidentified, and 259 clones were hypothetical, or unnamed proteins. Encoding 313 different sequences were identified as putative bio-defense genes or genes associated with immune response.

• Korean Journal of Ichthyology
• /
• v.19 no.4
• /
• pp.257-265
• /
• 2007

We constructed a cDNA library of testis from olive flounder (Paralichthys olivaceus) and a total of 248 expressed sequence tag (EST) clones were generated. In order to understand the molecular compositions of the olive flounder testis organs, the expression profiles of the identified clones in the cDNA library were analyzed. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 248 EST clones, 156 ESTs showed significant homology to previously described genes while 92 ESTs were unidentified or novel. Comparative analysis of the 156 identified ESTs showed that 6 (3.8%) clones were representing 5 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 (64.1%) clones representing 94 unique genes were identified as orthologs of known genes from other organisms, and orthologs were established for 50 (32.1%) clones representing 44 genes of known sequences with unknown functions. Furthermore, the testis library showed a more even distribution of cDNA clones with relatively fewer abundant clones that tend to contribute redundant clones in EST projects; thus, the testis library can supply more unique and novel cDNA sequences in olive flounder EST project.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2003.04a
• /
• pp.122-122
• /
• 2003

Expressed sequence tags (EST) are help to quickly identify functions of expressed genes and to understand the complexity of gene expression. In order to analyze gene expression of the leaf development in Panax ginseng, which is one of the most important medicinal plant, expressed sequence tags (EST) analysis was carried out. We constructed a cDNA library using the immature leaf of Chunpoong. Partial sequences were obtained from 3,170 clones. The ESTs could be clustered into 1,624 (56.1%) non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 1,137 groups show similarity to genes of known function. These ESTs clones were divided into sixteen categories depending upon gene function. Most abundant transcripts in immature ginseng leaf were photosynthesis related protein, such as chlorophyll a/b binding protein LHCII type I (128), chlorophyll a/b binding protein (53), ribulose-1,5-bisphosphate carboxylase (41), and photosystem I psaH (26). The EST data from immature leaf generated in this study is useful in dissecting gene expression in leaf organ of ginseng.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.7
• /
• pp.902-906
• /
• 2012

Expressed sequence tags (ESTs) from the Antarctic green algae Pyramimonas gelidicola were analyzed to obtain molecular information on cold acclimation of psychrophilic microorganisms. A total of 2,112 EST clones were sequenced, generating 222 contigs and 219 singletons, and 200 contigs and 391 singletons from control ($4^{\circ}C$) and cold-shock conditions ($-2^{\circ}C$), respectively. The complete EST sequences were deposited to the DDBJ EST database (http://www.ddbj.nig.ac.jp/index-e.html) and the nucleotide sequences reported in this study are available in the DDBJ/EMBL/GenBank. These EST databases of Antarctic green algae can be used in a wide range of studies on psychrophilic genes expressed by polar microorganisms.

• Journal of Aquaculture
• /
• v.16 no.1
• /
• pp.8-14
• /
• 2003

To generate expressed sequence tags for genomics research involving genetic linkage analysis, to examine gene expression profiles in muscles of channel catfish in a non-normalized muscle cDNA library, to compare gene expression in young and mature channel catfish muscles using the EST reagents and gene filters to demonstrate the feasibility of functional genomics research in small laboratories. 102 randomly picked cDNA clones were analyzed from the catfish muscle cDNA library. Of the sequences generated, 90.2% of ESTs was identified as known genes by identity comparisons. These 92 clones of known gene products represent transcriptional products of 24 genes. The 10 clones of unknown gene products represent 8 genes. The major transcripts (70.1% of the analyzed ESTs) in the catfish muscle are from many major genes involved in muscle contraction, relaxation, energy metabolism and calcium binding such as alpha actin, creatine kinase, parvalbumin, myosin, troponins, and tropomyosins. Gene expression of the unique ESTs was comparatively studied in the young and adult catfish muscles. Significant differences were observed for aldolase, myostatin, myosin light chain, parvalbumin, and an unknown gene. While myosin light chain and an unknown gene (CM 192) are down-regulated in the mature fish muscle, the aldolase, myostatin, and parvalbumin are significantly up-regulated in the mature fish muscle. Although the physiological significance of the changes in expression levels needs to be further addressed, this research demonstrates the feasibility and power of functional genomics in channel catfish. Channel catfish muscle gene expression profiles provide a valuable molecular muscle physiology blueprint for functional comparative genomics.

• Journal of fish pathology
• /
• v.22 no.3
• /
• pp.383-389
• /
• 2009

Analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics studies. As part of studies on the immune system of olive flounder, a total of 251 EST sequences from gill cDNA library were generated to identify and characterize important genes in the immune machanisms of olive flounder. Of the 251 clones, 126 clones (50.2%) were identified as orthologues of known genes from olive flounder and other organisms. Among the 126 EST clones, 16 clones (12.7%) were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 clones (79.4%) representing 103unique genes were identified as orthologs of known genes from other organisms. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immune related genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

• Journal of Life Science
• /
• v.10 no.2
• /
• pp.188-195
• /
• 2000

In order to understand molecular events during silk synthesis and provide genetic resources for molecular breeding, we had analyzed the cDNA library constructed from the posterior silkgland of Antheraea yamamai and partially sequenced 276 randomly selected genes from the cDNA library. Database comparisons of the expressed sequence tags (ESTs) revealed that 26 non-redundant clones showed a high similarity with previously identified genes. Among them, 17 clones exhibited a homology with previously identified insect genes and 9 clones were identical to genes that were previously identified from other organisms. A functional categorization showed that silk synthesis-defense- or stress-related genes, as well as genes involved in the metabolic pathways and in the transcriptional or translational apparatus are represented. In this report, the clone (AY479) which had high similarity with fibroin from A. pernyi was particularly analyzed in detail. The AY479 clone was carboxyl terminal region of fibroin. The 472 bp cDNA has 123 amino acids that shared 85% homology with the fibroin from A. pernyi and its deduced peptide had unique feature, that is, sites of alanine rich residues.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2003.04a
• /
• pp.123-123
• /
• 2003

In order to study gene expression transcribted during the embryo development, we constructed a cDNA library of embryogenic callus induced from cotylendon of Korean ginseng and generated expressed sequence tags (ESTs) of 3,359 clones randomly selected. The ESTs could be clustered into 1,910 (59.1%) non-redundant groups. Similarity search of the non-redundant ESTs against public non-redundant databases of both protein and DNA indicated that 2,217 groups show similarity to genes of known function. These ESTs clones were divided into eighteen categories depending upon gene function. Most abundant transcripts were ribosomal protein small subunit 28kDa(40), tumor-related protein(35), metallothionein (31), small heat-shock protein class 18.6K(24), and cyclophilin(20). There are no useful informations of gene expression during the embryo development in Korean ginseng. These results could help to understand the embryo development in Korean ginseng.

• Journal of fish pathology
• /
• v.23 no.2
• /
• pp.229-238
• /
• 2010

We constructed a rock bream (Oplegnathus fasciatus) liver cDNA library and a total of 1533 expressed sequence tag (EST) clones were generated. Gene annotation procedures and homology searches of the sequenced ESTs were analyzed using BLASTX. Of the 1533 EST clones, 1165 different ESTs showed significant homology to previously described genes while 368 ESTs were unidentified, hypothetical, or unnamed proteins. Encoding 106 different sequences were identified as putative bio-defense genes or genes associated with immune response.