• 분석과학
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• 제12권1호
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• pp.80-83
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• 1999

사람에게서 유래된 DNA시료의 X,Y 특이의 alphoid gene을 PCR법으로 분석하면 성별을 확인할 수 가 있다. PCR법으로 alphoid gene을 분석한바 매우 예민도가 높아 genomic DNA 약 60pg까지 성별을 분석할 수 있었다. 그리고 성별이 혼합되어 있는 DNA에서 female DNA의 1/10비 까지는 male DNA를 분석할 수 있었다. 따라서 이 결과는 혼합된 DNA에서 X,Y 특이의 alphoid gene을 분석하는데 기준으로 활용할 수가 있다.

• BMB Reports
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• 제37권4호
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• pp.474-479
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• 2004

The expressed sequence tag (EST) effort in Toxoplasma gondii has generated a substantial amount of gene information. To exploit this valuable resource, we chose to study tgd057, a novel gene identified by a large number of ESTs that otherwise show no significant match to known sequences in the database. Northern analysis showed that tgd057 is transcribed in this tachyzoite. The complete cDNA sequence of tgd057 is 1169 bp in length. Sequence analysis revealed that tgd057 possibly adopts two polyadenylation sites, utilizes the fourth in-frame ATG for translation initiation, and codes for a secretory protein. The longest open reading frame for the tgd057 gene was cloned and expressed as a recombinant protein (rd57) in Escherichia coli. Western analysis revealed that serum against rd57 recognized a molecule of ~21 kDa in the tachyzoite protein extract. This suggests that the tgd057 gene is expressed in vivo in the parasite.

• Asian-Australasian Journal of Animal Sciences
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• 제24권11호
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• pp.1529-1540
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• 2011

Silencing of a specific gene using RNAi (RNA interference) is a valuable tool for functional analysis of a target gene. However, information on RNAi for analysis of gene function in farm animals is relatively nil. In the present study, we have validated the interfering effects of siRNA (small interfering RNA) using both quantitative and qualitative gene silencing in buffalo granulosa cells. Qualitative gene knockdown was validated using a fluorescent vector, enhanced green fluorescence protein (EGFP) and fluorescently labeled siRNA (Cy3) duplex. While quantitatively, siRNA targeted against the luciferase and CYP19 mRNA was used to validate the technique. CYP19 gene, a candidate fertility gene, was selected as a model to demonstrate the technique optimization. However, to sustain the expression of CYP19 gene in culture conditions using serum is difficult because granulosa cells have the tendency to luteinize in presence of serum. Therefore, serum free culture conditions were optimized for transfection and were found to be more suitable for the maintenance of CYP19 gene transcripts in comparison to culture conditions with serum. Decline in fluorescence intensity of green fluorescent protein (EGFP) was observed following co-transfection with plasmid generating siRNA targeted against EGFP gene. Quantitative decrease in luminescence was seen when co-transfected with siRNA against the luciferase gene. A significant suppressive effect on the mRNA levels of CYP19 gene at 100 nM siRNA concentration was observed. Also, measurement of estradiol levels using ELISA (enzyme-linked immunosorbent assay) showed a significant decline in comparison to control. In conclusion, the present study validated gene silencing using RNAi in cultured buffalo granulosa cells which can be used as an effective tool for functional analysis of target genes.

• Journal of Plant Biotechnology
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• 제37권2호
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• pp.115-124
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• 2010

As the completion of genome sequencing, large collection of expression data and the great efforts in annotating plant genomes, the next challenge is to systematically assign functions to all predicted genes in the genome. Functional genome analysis of plants has entered the high-throughput stage. The generations and collections of mutants at the genome-wide level form technological platform of functional genomics. However, to identify the exact function of unknown genes it is necessary to understand each gene's role in the complex orchestration of all gene activities in the plant cell. Gene function analysis therefore necessitates the analysis of temporal and spatial gene expression patterns. The most conclusive information about changes in gene expression levels can be gained from analysis of the varying qualitative and quantitative changes of messenger RNAs, proteins and metabolites. New technologies have been developed to allow fast and highly parallel measurements of these constituents of the cell that make up gene activity. We have reviewed currently employed technologies to identify unknown functions of predicted genes including map-based cloning, insertional mutagenesis, reverse genetics, chemical mutagenesis, microarray analysis, FOX-hunting system, gene silencing mutagenesis, proteomics and chemical genomics. Recent improvements in technologies for functional genomics enable whole-genome functional analysis, and thus open new avenues for studies of the regulations and functions of unknown genes in plants.

• Genomics & Informatics
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• 제15권2호
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• pp.56-64
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• 2017

We have previously reported that NS-398, a cyclooxygenase-2 (COX-2)-selective inhibitor, inhibited replicative cellular senescence in human dermal fibroblasts and skin aging in hairless mice. In contrast, celecoxib, another COX-2-selective inhibitor, and aspirin, a non-selective COX inhibitor, accelerated the senescence and aging. To figure out causal factors for the senescence-modulating effect of the inhibitors, we here performed cDNA microarray experiment and subsequent Gene Set Enrichment Analysis. The data showed that several senescence-related gene sets were regulated by the inhibitor treatment. NS-398 up-regulated gene sets involved in the tumor necrosis factor ${\beta}$ receptor pathway and the fructose and mannose metabolism, whereas it down-regulated a gene set involved in protein secretion. Celecoxib up-regulated gene sets involved in G2M checkpoint and E2F targets. Aspirin up-regulated the gene set involved in protein secretion, and down-regulated gene sets involved in RNA transcription. These results suggest that COX inhibitors modulate cellular senescence by different mechanisms and will provide useful information to understand senescence-modulating mechanisms of COX inhibitors.

• 한국정보과학회논문지:소프트웨어및응용
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• 제37권1호
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• pp.1-8
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• 2010

최근 마이크로어레이 데이터를 기반으로 두 개의 샘플 그룹간에 유의한 발현 차이를 나타내는 생물학적 기능 그룹을 검출하기 위한 유전자 집합 분석(gene set analysis) 연구가 많은 주목을 받고 있다. 기존의 유의 유전자 검출 연구와는 달리, 유전자 집합 분석 연구는 유의한 유전자 집합과 이들의 기능적 특징을 함께 검출할 수 있다는 장점이 있다. 이러한 이유로 최근에는 PAGE, GSEA 등과 같은 다양한 통계적 방식의 유전자 집합 분석 방법들이 소개되고 있다. 특히, PAGE의 경우 두 샘플 그룹간의 유전자 발현 차이를 나타내는 스코어의 분포가 정규 분포임을 가정하는 모수적 접근 방식을 취하고 있다. 이러한 방법은 GSEA 등과 같은 비모수적 방식에 비해 계산량이 적고 성능이 비교적 우수한 장점이 있다. 하지만, PAGE에서 유전자 발현 차이를 정량화하기 위한 메트릭으로 사용하고 있는 AD(average difference)의 경우, 두 그룹간에 절대적 평균 발현 차이만을 고려하기 때문에 실제 유전자의 발현값 크기나 분산의 크기에 따른 상대적 중요성을 반영하지 못하는 문제가 있다. 본 논문에서는 이를 보완하기 위해 실제 유전자의 발현값 크기나 그룹 내 샘플들의 분산 정보 등을 스코어 계산에 함께 반영하는 WAD(weighted average difference), FC(Fisher's criterion), 그리고 Abs_SNR(Absolute value of signal-to-noise ratio)을 모수적 방식의 유전자 집합 분석에 적용하고 이에 따른 유의 유전자 집합 검출 결과를 실험을 통해 비교 분석하였다.

• 응용통계연구
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• 제31권2호
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• pp.241-251
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• 2018

마이크로어레이 자료의 유전자집합분석은 개별유전자분석에 비해 검정력도 높일 수 있고 결과 해석이 쉬워서 이에 대한 연구가 활발히 진행되어 왔다. 표현형에 따라 유의한 차이를 보이는 유전자집합의 검색은 검정통계량들이 유도된 배경에 따라 결과에 차이를 보이지만 대체적으로 t-통계량의 제곱합을 이용한 순열검정이 제일 무난한 방법으로 여겨진다. 그러나 유전자집합분석에서 다중검정은 필수이고 많은 집합들의 유의성에 변별력을 주기 위해서는 순열검정에서 생성하는 치환표본의 수가 많이 필요하고 시간이 오래 걸린다는 문제점이 있다. 순열검정을 대신할 모수적 방법들을 검토한 결과, 적률을 이용한 근사가 각 집합의 유의확률 계산시간도 훨씬 단축하며 순열검정에서 구한 유의확률과 크기와 순위가 거의 일치함을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권1호
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• pp.58-62
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• 2019

Objective: The oxidized low density lipoprotein receptor 1 (OLR1) gene plays an important role in the degradation of oxidized low-density lipoprotein and adipocyte proliferation in mammals. For this reason, we aimed at investigating the association of OLR1 gene polymorphisms with carcass quality traits in Chinese Qinchuan cattle. Methods: The single nucleotide polymorphism (SNP) was identified in the 3' untranslated region of bovine OLR1 gene by DNA sequencing. In addition, the haplotype frequency and linkage disequilibrium estimates of three SNPs were evaluated in 520 individuals. Results: Results indicated that the studied three SNPs were within the range of moderate genetic diversity (0.25< polymorphism information content<0.5). Haplotype analysis of three SNPs showed that ten different haplotypes were identified, but only five haplotypes were listed as those with a frequency of <0.05 were excluded. The Hap3 ($-G_1T_2C_3-$) had the highest haplotype frequency (42.10%). Linkage disequilibrium analysis showed that the three SNPs had a low linkage ($r^2<0.001$). The T10588C and C10647T were significantly associated with backfat thickness and intramuscular fat content in Qinchuan cattle. Conclusion: Based on our results, we believe that the OLR1 gene could be a strong candidate gene for influencing carcass quality traits in Qinchuan cattle.

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.115-121
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• 1992

In order to broaden the spectrum of substrate utilization of a Gram negative bacterium Zymomonas mobilis which has a great potential as an industrial ethanol producing microorganism, cloning of $\alpha$-amylase gene into Z. mobilis ZM4 was tried. The $\alpha$-amylase gene was isolated from Bacillus stearothermophilus. By Southern blot analysis, it was proven that the $\alpha$-amylase gene fragment was originated from a naturally occuring plasmid of B. stearothermophilus ATCC 31195. To place $\alpha$-amylase gene under the control of Z. mobilis promoter, two different Z. mobilis expression vectors, pZA26 and pLOI204, were used. The truncated $\alpha$-amylase gene was then introduced into these vectors. Both qualitative and quantitative activities of $\alpha$-amylase were observed in Z. mobilis cells harboring these plasmids with the $\alpha$-amylase gene inserted. Gas chromatographic analysis of ethanol showed that one of the Z. mobilis transconjugants was capable of producing 67 mM ethanol from rich medium(RM) containing 5% soluble starch as a sole carbon source.

• BMB Reports
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• 제47권6호
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• pp.348-353
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• 2014

The speed of sound (SOS) value is an indicator of bone mineral density (BMD). Previous genome-wide association (GWA) studies have identified a number of genes, whose variations may affect BMD levels. However, their biological implications have been elusive. We re-analyzed the GWA study dataset for the SOS values in skeletal sites of 4,659 Korean women, using a gene-set analysis software, GSA-SNP. We identified 10 common representative GO terms, and 17 candidate genes between these two traits (PGS < 0.05). Implication of these GO terms and genes in the bone mechanism is well supported by the literature survey. Interestingly, the significance levels of some member genes were inversely related, in several gene-sets that were shared between two skeletal sites. This implies that biological process, rather than SNP or gene, is the substantial unit of genetic association for SOS in bone. In conclusion, our findings may provide new insights into the biological mechanisms for BMD.