• Food Science of Animal Resources
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• v.28 no.3
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• pp.276-282
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• 2008

It is estimated that over 80% of deer antlers produced in the world are consumed in Korea. However, mislabeling or fraudulent replacement of costly antlers with cheaper ones is one of the most common problems in the domestic antler market. Therefore, there is a great need for the development of technology to identify species of antlers. This study was carried out to develop an accurate and reliable method for the identification and authentication of species or subspecies of antlers using DNA sequence analysis and comparison of mitochondrial cytochrome band D-loop region genes among antlers of five deer species, Cervus elaphus sibericus, Cervus elaphus canadensis, Cervus nippon, Cervus elaphus bactrianus and Rangifer tarandus. A variable region of cytochrome band D-loop genes was amplified using PCR with specifically designed primers and sequenced directly. The cytochrome band D-loop region genes showed different DNA sequences between the species of antlers and thus it is possible to differentiate between species on the basis of sequence variation. To distinguish between reindeer (Rangifer tarandus) antlers and other deer antlers, PCR amplicons of the cytochrome b gene were digested with the restriction enzymes NlaIV and TaqI, respectively, which generates a species-specific DNA profile of the reindeer. In addition, samples of 32 sliced antlers labeled Cervus elaphus sibericus from commercial markets were collected randomly and the mt DNA D-loop region of these antler samples was sequenced. Among the antler samples investigated, only 62.5% were from Cervus elaphus sibericus, and others were from Cervus elaphus bactrianus (25.0%), elk (Cervus elaphus canadensis) and reindeer (Rangifer tarandus). Our results suggest that DNA sequencing of mt DNA and PCR-RFLP methods using NlaIV and TaqI enzymes are useful for the identification and discrimination of deer antler species by routine analysis.

• Korean Journal of Plant Resources
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• v.30 no.5
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• pp.522-534
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• 2017

This study was investigated genetic variation in 24 Taraxacum accessions from various regions in South Korea based on the sequences of two chloroplast DNA (cpDNA) regions (trnL-trnF and rps16-trnK). T. mongolicum, T. officinale, and T. laevigatum were triploid, and T. coreanum and T. coreanum var. flavescens were tetraploid. The trnL-trnF region in native Korean dandelions (T. mongolicum, T. coreanum, and T. coreanum var. flavescens) were ranged from 931 to 935 bp in length, and that of naturalized dandelions were ranged from 910 bp (T. officinale) to 975 bp (T. laevigatum) in length. The rps16-trnK region in T. mongolicum, T. coreanum, T. coreanum var. flavescens, T. officinale, and T. laevigatum was 882-883 bp, 875-881 bp, 878-883 bp, 874-876 bp, and 847-876 bp, respectively, in length. The sequence similarity matrix of the trnL-trnF region ranged from 0.860 to 1.00 with an average of 0.949, and that of the rps16-trnK region ranged from 0.919 to 1.000 with an average of 0.967. According to the phylogenetic analysis, the Korean native taxa and naturalized taxa were divided independent clade in two cpDNA region. T. coreanum var. flavescens clustered only with T. coreanum, and there were no significant differences in their nucleotide sequences. The finding that two accessions (T. coreanum; Jogesan, T. mongolicum; Gangyang) had a high level of genetic variation suggests their utility for breeding materials.

• Bioinformatics and Biosystems
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• v.1 no.2
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• pp.99-102
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• 2006

The Basic Local Alignment Search Tool (BLAST) is one of the most established software in bioinformatics research and it compares a query sequence against the libraries of known sequences in order to investigate sequence similarity. Expressed Sequence Tags (ESTs) are single-pass sequence reads from mRNA (or cDNA) and represent the expression for a given cDNA library and the snapshot of genes expressed in a given tissue and/or at a given developmental stage. Therefore, ESTs can be very valuable information for functional genomics and bioinformatics researches. Although major bio database (DB) websites including NCBI are providing BLAST services and EST data, local DB and search system is demanding for better performance and security issue. Here we present animal EST DBs and local BLAST search system. The animal ESTs DB in NCBI Genbank were divided by animal species using the Perl script we developed. and we also built the new extended DB search systems fur the new data (Local Animal BLAST Search System: http://bioinfo.kohost.net), which was constructed on the high-capacity PC Cluster system fur the best performance. The new local DB contains 650,046 sequences for Bos taurus(cattle), 368,120 sequences for Sus scrofa (pig), 693,005 sequences for Gallus gallus (fowl), respectively.

• The Korean Journal of Malacology
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• v.30 no.2
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• pp.155-163
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• 2014

Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.909-917
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• 1998

Six different sequences types(A through E and H) and a subtype(Bl) of the small subunit ribosomal RNA(SSUrRNA) gene were found in bovine Theileria isolates from different areas of Korea and Japan. The sequences were aligned and three hypervariable regions were observed in the nucleotide position ranges 212~231, 261~270 and 632~690. Five of the Theileria isolates yielded sequence type A; these were the field isolates KCB, KCN, and KCJ, and the laboratory stock KLS, all from Korea, and a single isolate from Japan (JHS). This sequence type is identical to the SSUrRNA gene sequence listed for Theileria buffeli (GenBank Accession No. Z15106) from Marula, Kenya. The Korean field isolate KKB yielded only a single sequence type (B), but multiple sequence types were found in some isolates. For example, KCB and JHS isolates yielded both types A and B ; isolate KKW showed types B and H; isolate KCN showed types A, C, and D ; and isolate KCJ showed types A, B, E, and a subtype B1. Finding of the multiple sequences SSUrRNA gene sequences suggests that bovine Theileria isolates from both Korea and Japan may consist of mixed populations.

• Journal of Life Science
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• v.22 no.2
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• pp.142-147
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• 2012

The sporulation-specific glucoamylase (SGA) of Saccharomyces diastaticus is known to be produced in the cytoplasm during sporulation. For the purpose of proving that SGA has secretory potential, we constructed a hybrid plasmid, pYESC25, containing the promoter and the putative signal sequence of the SGA fused in frame to the endo-1,4-${\beta}$-D-glucanase (CMCase) gene of Bacillus subtilis without its own signal sequence. The recipient yeast strain of S. diastaticus YIY345 was transformed with the hybrid plasmid. CMCase secretion from S. diastaticus harboring pYESC25 into culture medium was confirmed by the formation of yellowish halos around transformants after staining with Congo red on a CMC agar plate. The transformant culture was fractionated to the extracellular, periplasmic, and intracellular fraction, followed by the measurement of CMCase activity. About 63% and 13% enzyme activity were detected in the culture supernatant (extracellular fraction) and periplasmic fraction, respectively. Furthermore, ConA-Sepharose chromatography, native gel electrophoresis, and activity staining revealed that CMCase produced in yeast was glycosylated and its molecular weight was larger than that of the unglycosylated form from B. subtilis. Taking these findings together, SGA has the potential of secretion to culture medium, and the putative signal sequence of SGA can efficiently direct bacterial CMCase to the yeast secretion pathway.

• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.274-285
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• 2007

The subfamily Chrominae of damselfishes (Teleostei: Pomacentridae) includes the genus Chromis and Dascyllus. They are found throughout the tropical oceans and form a major component of coral reef communities. There are 5 species of the Chrominae currently recognized in Korea. This study was conducted to infer phylogenetic position of two Korean Chromis species and one Dascyllus species within general category of their each genus in worldwide level. This study also includes one species of Japanese Dascyllus. In the phylogenetic analysis, the Japanese D. aruanus grouped with D. aruanus previously reported from French Polynesia. Korean Chromis fumea grouped with Australian C. nitida and the p-distance value between the two species is relatively very low (0.047). Korean C. notatus grouped together with C. flavomaculata (New Caledonia). In the sequence analysis of some Korean and Japanese damselfishes, there was no sequence variation between D. melanurus (Jeju, Korea) and D. melanurus (Indo-Pacific), but the sequences of the two populations were different in only one nucleotide sites from that of D. melanurus in Indonesian Archipelago. The sequences of Dascyllus aruanus (Japan) were different in two nucleotide sites from it in French Polynesia. There were high difference between the sequences of two Korean species, Chromis fumea and Korean C. notatus. The variations among mitochondrial cytochrome b sequences indicate that the gene sequence could be used as DNA barcode for identification of local populations of D. aruaus and D. melanurus as well as species level.

• Journal of Animal Science and Technology
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• v.51 no.1
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• pp.1-8
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• 2009

We generated 57,598 expressed sequence tags (ESTs) from 3 cDNA libraries of Hanwooo (Korean Cattle), fat, loin, liver. Liver, intermuscular fat and longissimus dorsi tissues were obtained from a 24-month-old Hanwoo steer immediately after slaughter. cDNA library was constructed according to the oligocapped method. The EST data were clustered and assembled into unique sequences, 4,759 contigs and 7,587 singletons. To carry out functional analysis, Gene Ontology annotation and identification of significant leaf nodes were performed that were detected by searching significant p-values from $2^{nd}$ level GO terms to leaf nodes using Bonferroni correction. We found that 13, 26 and 8 significant leaf nodes are unique in the transcripts according to 3 GO categories, molecular function, biological process and cellular component. Also digital gene expression profiling using the Audic's test was performed and tissue specific genes were detected in the above 3 libraries.

• Journal of Life Science
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• v.17 no.9 s.89
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• pp.1248-1254
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• 2007

The secretory overexpression of Clostridium thermocellum endoglucanase A gene (celA) was examined in Saccharomyces cerevisiae using Kluyveromyces marxianus exoinulinase (INU1) signal sequence and GAL10 promoter. The two plasmids, pYEG-CT1 with its own signal sequence, and pYInu-CT1 with INU1 signal sequence were introduced to S. cerevisiae SEY2102 and S. cerevisiae 2805 host strains, respectively, and then each transformant was selected on the synthetic defined media lacking uracil. The expression level and secretion efficiency of endoglucanase A was increased by $18{\sim}22%$ and 11%, respectively, by INU1 signal sequence over celA signal sequence. By considering the high level of expression (361 unit/I), plasmid stability (89%), and secretion efficiency (70%), S. cerevisiae 2805 harboring plasmid pYInu-CT1 was selected as the opti-mal host vector system for the production of cellulose-degrading enzyme and recombinant yeast probiotic. The total expression and secretion efficiency of endoglucanase A was 418 unit/l and 73%, respectively, in the batch fermentation of S. cerevisiae 2805/pYlnu-CT1 on galactose medium. The mo-lecular weight of secreted endoglucanase A was found to be greater than 100 kDa, presumably due to the N-linked glycosylation.

• Korean Journal of Microbiology
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• v.39 no.1
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• pp.1-7
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• 2003

The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).