• 한국임상수의학회지
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• 제33권6호
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• pp.346-350
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• 2016

Two 1-year old calves of Korean Native cattle (Hanwoo) presented cutaneous papillomas on the face and neck. Type 2 bovine papillomavirus (BPV-2) was identified in the cutaneous papillomas based on BPV-specific PCR and subsequent DNA sequencing analysis results. Using DNA samples extracted from two affected calves and unaffected animals reared in the same stable, BPV-2 was not only detected in the cutaneous papillomas of affected animals based on BPV-specific PCR analysis, but also detected in normal skins, hairs, and their environments based on nested PCR analysis. BPV-2 was also detected in DNA samples isolated from animals and environments of that distinct stable with affected calves. However, no BPV-2 was detected in the drinking water of both stables (infected and unaffected). These findings concluded that BPV-2 was transmitted by direct or indirect contact, not by drinking water. This is the first report to show molecular evidence of BPV-2 infection. Rapid and precise molecular identification can be used to screen BPV-2 in cattle farms to understand the biological roles of BPV in animal diseases.

• 동의생리병리학회지
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• 제19권6호
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• pp.1534-1540
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• 2005

In order to testify the urate lowering effects of Daihwangmudan-tang(DMT), ICR mice were injected monosodium urate into the abdominal cavity and then DMT was administered on 2 and 4 days after Injection. Uric acid and triglyceride were measured as hematological indices of gout, and some genes related with this change were identified by ACP based GeneFishing PCR method and direct sequencing. From this experiment, DMT highly decreased the blood levels of uric acid and significantly suppressed and lowered the acute increment of triglyceride level. There were 11 differentially expressed genes(DEG) having relations with positive actions of DMT, and 4 major genes in the middle of DEGs were sequenced; Mfap 2, jagged 2, Hsd17b7, DkkI-1, These genes were supposed that several mechanisms through interleukin 1 and T-cell anergy, LDL cholesterol metabolism, wnt pathway would be related with the anti-inflammation effect against gout.

• 분석과학
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• 제15권3호
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• pp.190-195
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• 2002

본 연구에서는 ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC)방식을 이용한 denaturing high performance liquid chromatography (DHPLC)방법을 사용하여 기관지 천식을 조절하는 ${\beta}_2$-교감신경수용체 유전자의 돌연변이를 검출하였다. 50명의 천식 환자의 혈액에서 genomic DNA를 추출하여 중합효소연쇄반응 (polymerase chain reaction, PCR)을 이용해 증폭하고, 그 산물을 DHPLC로 분석한 결과, 50개의 시료 가운데 15개 (30%)의 돌연변이를 검출하였다. 최종적으로 유전자 염기서열결정법을 통해 DHPLC의 돌연변이 검출률이 정확함을 확인하였다.

• Animal cells and systems
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• 제16권6호
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• pp.498-505
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• 2012

This study uses the mitochondrial DNA control region to identify the genetic diversity and population structure of the marbled soles (Pleuronectes yokohamae) that inhabit Jinhae Bay and Yokji Island in the nearby sea and the adjacent waters of Namhae, Hansan Island, and Jaran Bay. Direct sequencing of the PCR products revealed 379 bp sequences with 83 variable nucleotide sites, defining a total of 91 haplotypes. The haplotype diversity was high, ranging from $0.917{\pm}0.031$ to $0.983{\pm}0.008$, and nucleotide diversity ranged from $0.015{\pm}0.008$ to $0.024{\pm}0.012$. In addition, 48 haplotypes (52.7%) were unique. Pairwise $F_{ST}$ values were very low, with the maximum value occurring between PYH (Hansan Island) and PJI (Jinhae Bay) ($F_{ST}$ = 0.011). Therefore, no significant genetic differentiation was evident between any pair of sampling localities.

• Mycobiology
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• 제31권4호
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• pp.196-199
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• 2003

A very reliable and specific method for the identification of fungi in ectotrophic mycorrhizal symbiosis was developed using a specific PCR assay based on the amplification of the ITS1 region. To obtain specific data, an ITS-diagnostic assay was carried out that reveals genera and species specific sequences. Here, an application of one method is presented, which covers the identification of pure mycelia, basidiocarps as well as mixed samples such as ectomycorrhizal roots that were mingled with remains of the host plant. For this purpose a protocol was established that allowed the extraction of DNA from single mycorrhizal roots. In order to perform a specific ITS analysis we generated a new ITS-primer(ITS8) by a multiple alignment of five different genera and species of mycorrhizal fungi. The utilization of ITS1 and ITS8 resulted in specific PCR amplicons, which were characterized by sequencing without purification steps, even when the template DNA was associated with roots.

• Clinical and Experimental Pediatrics
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• 제60권10호
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• pp.327-332
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• 2017

Purpose: Short stature affects approximately 2%-3% of children, representing one of the most frequent disorders for which clinical attention is sought during childhood. Despite assumed genetic heterogeneity, mutations or deletions in the short stature homeobox-containing gene (SHOX ) are frequently detected in subjects with short stature. Idiopathic short stature (ISS) refers to patients with short stature for various unknown reasons. The goal of this study was to screen all the exons of SHOX to identify related mutations. Methods: We screened all the exons of SHOX for mutations analysis in 105 ISS children patients (57 girls and 48 boys) living in Taif governorate, KSA using a direct DNA sequencing method. Height, arm span, and sitting height were recorded, and subischial leg length was calculated. Results: A total of 30 of 105 ISS patients (28%) contained six polymorphic variants in exons 1, 2, 4, and 6. One mutation was found in the DNA domain binding region of exon 4. Three of these polymorphic variants were novel, while the others were reported previously. There were no significant differences in anthropometric measures in ISS patients with and without identifiable polymorphic variants in SHOX. Conclusion: In Saudi Arabia ISS patients, rather than SHOX, it is possible that new genes are involved in longitudinal growth. Additional molecular analysis is required to diagnose and understand the etiology of this disease.

• Journal of Microbiology and Biotechnology
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• 제20권5호
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• pp.853-861
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• 2010

High substrate costs decrease the profitability of polyhydroxyalkanoates (PHAs) production, and thus low-cost carbon substrates coming from agricultural and industrial residuals are tested for the production of these biopolymers. Among them, crude glycerol, formed as a by-product during biodiesel production, seems to be the most promising source of carbon. The object of this study was to characterize the mixed population responsible for the conversion of crude glycerol into PHAs by cultivation-dependent and -independent methods. Enrichment of the microbial community was monitored by applying the Ribosomal Intergenic Spacer Analysis (RISA), and the identification of community members was based on 16S rRNA gene sequencing of cultivable species. Molecular analysis revealed that mixed populations consisted of microorganisms affiliated with four bacterial lineages: ${\alpha}$, ${\gamma}$-Proteobacteria, Actinobacteria, and Bacteroides. Among these, three Pseudomonas strains and Rhodobacter sp. possessed genes coding for polyhydroxyalkanoates synthase. Comparative analysis revealed that most of the microorganisms detected by direct molecular analysis were obtained by the traditional culturing method.

• 대한의생명과학회지
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• 제17권1호
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• pp.27-37
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• 2011

Hepatitis A virus (HAV) is a causative agent of triggering acute hepatitis which is transmitted by person-to-person contact and or fecal-oral route. In previous studies, most hepatitis A virus (HAV) isolates had been genotype IA in Korea. Recently, a small number of different genotypes were reported with an upsurge of acute hepatitis by HAV. Therefore, the distribution of HAV genotypes was investigated. RNA was extracted from anti-HAV IgM positive sera which were collected from February to August 2009, at a tertiary care hospital in eastern Jeonnam, Korea. Nested reverse transcription PCR and direct sequencing for VP1/P2A region of the HAV were performed. A total of 365 cases with suspected acute hepatitis were tested for anti-HAV IgM and positive results were obtained in 24 sera (9.0%), which were collected 2 to 15 days (median, 7 days) after the onset of symptoms. Of the 24 seropositive samples, 14 (58.3%) samples were positive for HAV RNA, among which 4 isolates (28.6%) were genotype IA and the other 10 (71.4%) were genotype IIIA. Both IA and IIIA genotypes were isolated from 5~6 neighboring administrative districts throughout the year without geographic or seasonal restrictions. HAV genotypes (IA and IIIA) were observed from the eastern Jeonnam for the studied.

• 한국동물학회지
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• 제35권1호
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• pp.80-86
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• 1992

The nucleotide sequences of 185 rRNAs of the five Korean decapods were partially determined by the direct sequencing method using the reverse transcriptase. ne average GC content of five species was 51.1% which is higher than that of yeast(45.0%) and lower than those of frog (53.0%) and rat (55.6%). This result follows the general patterns of the GC content in the nucleotides of the nucleic acid shown among the various phylogenetic groups. The average ratio of transrional/transversional nucleotide substitution of pairwise comparison among six species (including Anemia salina) was 1.200 $\pm$ 0.310 when whole region alas examined. However, the ratio showed some differences when the conservative regions and variable regions frere separatelv examined. The molecular phylogenies of the five species were constructed by using two different tree making methods. In general the results support the previously reported molecular phylogeny of the decapod crustaceans. However, our results indicate thats in the analysis of the sequence dat3, the UPGMA clustering method of the distance matrix method should be carefully employed after considering the rate of nucneotide substitution in the different regions of the molecule.

• KSBB Journal
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• 제26권2호
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• pp.131-138
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• 2011

Some computational approaches are needed for clarifying RNAi sequences, because it takes much time and endeavor that almost of RNAi sequences are verified by experimental data. Incorrectness of RNAi mechanism and other unaware factors in organism system are frequently faced with questions regarding potential use of RNAi as therapeutic applications. Our massive parallelized pair alignment scoring between dsRNA in Genebank and expressed sequence tags (ESTs) in Caenorhabditis elegans Genome Sequencing Projects revealed that this provides a useful tool for the prediction of RNAi induced cosuppression details for practical use. This pair alignment scoring method using high performance computing exhibited some possibility that numerous unwanted gene silencing and cosuppression exist even at high matching scores each other. The classifying the relative higher matching score of them based on GO (Gene Ontology) system could present mapping dsRNA of C. elegans and functional roles in an applied system. Our prediction also exhibited that more than 78% of the predicted co-suppressible genes are located in the ribosomal spot of C. elegans.