• Title/Summary/Keyword: Candidate gene approach

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Isolation of Deletion Mutants by Reverse Genetics in Caenorhabditis elegans

  • Park, Byung-Jae;Lee, Jin ll;Lee, Jiyeon;Kim, Sunja;Choi, Kyu Yeong;Park, Chul-Seung;Ahn, Joohong
    • Animal cells and systems
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    • v.5 no.1
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    • pp.65-69
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    • 2001
  • Obtaining mutant animals is important for studying the function of a particular gene. A chemical mutagenesis was first carried out to generate mutations in C. elegans. In this study, we used ultraviolet-activated 4,5',8-trimethylpsoralen to induce small deletion mutations. A library of mutagenized worms was prepared for recovery of candidate animals and stored at $15^{\circ}C$ during screening instead of being made into a frozen stock library. In order to isolate deletion mutations in target genes, a polymerase chain reaction (PCR)-based screening method was used. As a result, two independent mutants with deletions of approximately 1.0 kb and 1.3 kb were isolated. This modified and improved reverse genetic approach was proven to be effective and practical for isolating mutant animals to study gene function at the organismal level.

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Association Analysis of Reactive Oxygen Species-Hypertension Genes Discovered by Literature Mining

  • Lim, Ji Eun;Hong, Kyung-Won;Jin, Hyun-Seok;Oh, Bermseok
    • Genomics & Informatics
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    • v.10 no.4
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    • pp.244-248
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    • 2012
  • Oxidative stress, which results in an excessive product of reactive oxygen species (ROS), is one of the fundamental mechanisms of the development of hypertension. In the vascular system, ROS have physical and pathophysiological roles in vascular remodeling and endothelial dysfunction. In this study, ROS-hypertension-related genes were collected by the biological literature-mining tools, such as SciMiner and gene2pubmed, in order to identify the genes that would cause hypertension through ROS. Further, single nucleotide polymorphisms (SNPs) located within these gene regions were examined statistically for their association with hypertension in 6,419 Korean individuals, and pathway enrichment analysis using the associated genes was performed. The 2,945 SNPs of 237 ROS-hypertension genes were analyzed, and 68 genes were significantly associated with hypertension (p < 0.05). The most significant SNP was rs2889611 within MAPK8 (p = $2.70{\times}10^{-5}$; odds ratio, 0.82; confidence interval, 0.75 to 0.90). This study demonstrates that a text mining approach combined with association analysis may be useful to identify the candidate genes that cause hypertension through ROS or oxidative stress.

Proteomic Comparison of Gibberella moniliformis in Limited-Nitrogen (Fumonisin-Inducing) and Excess-Nitrogen (Fumonisin-Repressing) Conditions

  • Choi, Yoon-E;Butchko, Robert A.E.;Shim, Won-Bo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.780-787
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    • 2012
  • The maize pathogen Gibberella moniliformis produces fumonisins, a group of mycotoxins associated with several disorders in animals and humans, including cancer. The current focus of our research is to understand the regulatory mechanisms involved in fumonisin biosynthesis. In this study, we employed a proteomics approach to identify novel genes involved in the fumonisin biosynthesis under nitrogen stress. The combination of genome sequence, mutant strains, EST database, microarrays, and proteomics offers an opportunity to advance our understanding of this process. We investigated the response of the G. moniliformis proteome in limited nitrogen (N0, fumonisin-inducing) and excess nitrogen (N+, fumonisin-repressing) conditions by one- and two-dimensional electrophoresis. We selected 11 differentially expressed proteins, six from limited nitrogen conditions and five from excess nitrogen conditions, and determined the sequences by peptide mass fingerprinting and MS/MS spectrophotometry. Subsequently, we identified the EST sequences corresponding to the proteins and studied their expression profiles in different culture conditions. Through the comparative analysis of gene and protein expression data, we identified three candidate genes for functional analysis and our results provided valuable clues regarding the regulatory mechanisms of fumonisin biosynthesis.

Genetic Polymorphism of Interleukin 10 Gene and Sasang Constitution in Bell's Palsy Patients

  • Kim, Jong-Won;Seo, Jung-Chul;Jung, Tae-Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.515-519
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    • 2005
  • We hypothesized that the IL10 gene is important candidate in the development of Bell's palsy and specific genotypic and allelic variations should be associated with Bell's palsy in the Korean population. In this study, we assessed the SNP (single-nucleotide polymorphism) of IL10 in patients with Bell's palsy. 62 patients with Bell's palsy were selected from the subjects who visited for the Bell's palsy service of the department of acupuncture & moxibustion, college of Oriental Medicine, Daegu Haany University from May 2002 to May 2003. Pyrosequencing was performed for genetic analyses. There was no statistically significant genotypic distribution difference between control and Bell's palsy group And there was not statistically significant allelic frequency difference between control and Bell's palsy group. In this study the IL10 genotypemight not be the risk factor of Bell's palsy patients in Korean. studies will be necessary for the exact genetic markers. Establishment of more systemic approach and high quality of prospective cohorts will be necessary for the good prediction of genetic markers.

Pathway Analysis in HEK 293T Cells Overexpressing HIV-1 Tat and Nucleocapsid

  • Lee, Min-Joo;Park, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1103-1108
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    • 2009
  • The human immunodeficiency virus (HIV)-l protein Tat acts as a transcription transactivator that stimulates expression of the infected viral genome. It is released from infected cells and can similarly affect neighboring cells. The nucleocapsid is an important protein that has a related significant role in early mRNA expression, and which contributes to the rapid viral replication that occurs during HIV-1 infection. To investigate the interaction between the Tat and nucleocapsid proteins, we utilized cDNA micro arrays using pTat and flag NC cotransfection in HEK 293T cells and reverse transcription-polymerase chain reaction to validate the micro array data. Four upregulated genes and nine downregulated genes were selected as candidate genes. Gene ontology analysis was conducted to define the biological process of the input genes. A proteomic approach using PathwayStudio determined the relationship between Tat and nucleocapsid; two automatically built pathways represented the interactions between the upregulated and downregulated genes. The results indicate that the up- and downregulated genes regulate HIV-1 replication and proliferation, and viral entry.

Characterization of Diesel Degrading Enterobacter cancerogenus DA1 from Contaminated Soil

  • Kim, Sang-Jun;Joo, Gil-Jae
    • Korean Journal of Environmental Biology
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    • v.36 no.2
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    • pp.190-198
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    • 2018
  • The petroleum industry is an important part of the world economy. However, the massive exposure of petroleum in nature is a major cause of environmental pollution. Therefore, the microbial mediated biodegradation of petroleum residues is an emerging scientific approach used to resolve these problem. Through the screening of diesel contaminated soil we isolated a rapid phenanthrene and a diesel degrading bacterium identified as Enterobacter cancerogenus DA1 strain through 16S rRNA gene sequence analysis. The strain was registered in NCBI with an accession number MG270576. The optimal growth condition of the DA1 strain was determined at pH 8 and $35^{\circ}C$, and the highest degradation rate of the diesel was achieved at this condition. At the optimal condition, growth of the strain on the medium containing 0.05% phenanthrene and 0.1% of diesel-fuel was highest at 45 h and 60 h respectively after the incubation period. Biofilm formation was found significantly higher at $35^{\circ}C$ as compared to $30^{\circ}C$ and $40^{\circ}C$. Likewise, the lipase activity was found significantly higher at 48 h after the incubation compared to 24 h and 72 h. These results suggest that the Enterobacter cancerogenus DA1 could be an efficient candidate, for application through ecofriendly scientific approach, for the biodegradation of petroleum products like diesel.

The Effects of Estrogen Receptor Locus on Reproductive Tracts Components and Performance Traits in Large White×Meishan F2 Offspring

  • Li, Fenge;Lei, Minggang;Zheng, Rong;Zuo, Bo;Jiang, Siwen;Deng, Changyan;Xiong, Yuanzhu
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.9
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    • pp.1223-1226
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    • 2004
  • Previously candidate gene approach revealed estrogen receptor (ESR) locus was associated with increased litter size. In this study, PvuII polymorphisms of ESR gene was detected by PCR-RFLP, and ESR locus was evaluated for its association with reproductive tracts components in the Large $White{\times}Meishan$ ($LW{\times}M$) F2 offspring. Ninety seven gilts with reproductive tracts components records and 136 offspring with performance traits records were genotyped and the results were used to estimate allele substitution effects. The results showed that two alleles (A and B) were identified, and 121 bp fragments were observed for the AA genotype and 65 bp and 56 bp fragments for the BB genotype; the length of uterine body (LUB) of BB gilts were significantly shorter than AA gilts', the additive effect was -1.762 cm; the uterine weight (UW) of AB gilts were significantly lighter than AA gilts' with the additive effect -18.058 g; no significant associations of ESR alleles with ovulation rate (OR), length of uterine horn (LUH), length of uterine cervix (LUC), weight of two ovaries (OW), volume of uterine lumen (VUL), length of oviduct (LO) were observed. BB genotypes gilts need significantly less days to 100 kg ($D_{100kg}$) than AA genotypes (p<0.01), the additive effect was per copy of B allele. Allele B is also favorable for average daily gain (ADG), with additive effect 0.015 kg/d (p<0.05). There was no difference between genotypes for backfat thickness at the 13th rib (SF13), loin meat height (ELMH), and loin meat percentage was estimated (ELMP), individual birth weight (IBW) and teat number (TN).

Systems Pharmacological Approach to Identification of Schizonepeta teunifolia Extract via Active Ingredients Analysis and Cytotoxicity Effect on A549 Cell Lines (형개 추출물의 시스템 약리학적 분석과 비소세포폐암세포에 대한 증식 억제효과)

  • Ga Ram Yang;Ji Eun Choo;Youn Sook Kim;Won Gun Ahn
    • Korean Journal of Acupuncture
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    • v.41 no.1
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    • pp.7-15
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    • 2024
  • Objectives : This study aimed to predict the effectiveness and potential of Schizonepeta tenuifolia as an anticancer treatment for non-small cell lung cancer through network-based pharmacology and cellular experiment. Methods : To identify the major bioactive compounds in Schizonepeta tenuifolia, we used the Traditional Chinese Medicine Systems. The target genes for the cancer treatment were selected using the UniProt database and the networked using Cytoscape. We performed functional enrichment analysis based on the Gene Ontology Biological Process and Kyoto Encyclopedia of Genes and Genomes Pathways to predict the mechanisms. To investigate the effect of Schizonepeta tenuifolia on lung cancer cell growth, we treated A549 cells, a lung cancer cell line, with different concentrations of the drug and used the MTT assay for cell viability. Results : Research has shown that the most effective mechanism of active compounds from Schizonepeta tenuifolia is through the pathway of cancer. The results of the network pharmacology analysis indicate that Schizonepeta tenuifolia has potential medicinal value as an adjuvant in anticancer treatment. The concentration-dependent inhibition of cell viability was observed on A549 cells. Furthermore, synergistic anticancer activity with Doxorubicin was also observed. Conclusions : Through a network pharmacological approach, Schizonepeta tenuifolia was predicted to have potential as an anticancer agent, and its efficacy was experimentally demonstrated using A549 cells. These findings suggest that Schizonepeta tenuifolia is a promising candidate for future research.

Genome-wide association study for the free amino acid and nucleotide components of breast meat in an F2 crossbred chicken population

  • Minjun Kim;Eunjin Cho;Jean Pierre Munyaneza;Thisarani Kalhari Ediriweera;Jihye Cha;Daehyeok Jin;Sunghyun Cho;Jun Heon Lee
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.57-68
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    • 2023
  • Flavor is an important sensory trait of chicken meat. The free amino acid (FAA) and nucleotide (NT) components of meat are major factors affecting meat flavor during the cooking process. As a genetic approach to improve meat flavor, we performed a genome-wide association study (GWAS) to identify the potential candidate genes related to the FAA and NT components of chicken breast meat. Measurements of FAA and NT components were recorded at the age of 10 weeks from 764 and 767 birds, respectively, using a White leghorn and Yeonsan ogye crossbred F2 chicken population. For genotyping, we used 60K Illumina single-nucleotide polymorphism (SNP) chips. We found a total of nine significant SNPs for five FAA traits (arginine, glycine, lysine, threonine content, and the essential FAAs and one NT trait (inosine content), and six significant genomic regions were identified, including three regions shared among the essential FAAs, arginine, and inosine content traits. A list of potential candidate genes in significant genomic regions was detected, including the KCNRG, KCNIP4, HOXA3, THSD7B, and MMUT genes. The essential FAAs had significant gene regions the same as arginine. The genes related to arginine content were involved in nitric oxide metabolism, while the inosine content was possibly affected by insulin activity. Moreover, the threonine content could be related to methylmalonyl-CoA mutase. The genes and SNPs identified in this study might be useful markers in chicken selection and breeding for chicken meat flavor.

Detection of Gene Interactions based on Syntactic Relations (구문관계에 기반한 유전자 상호작용 인식)

  • Kim, Mi-Young
    • The KIPS Transactions:PartB
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    • v.14B no.5
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    • pp.383-390
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    • 2007
  • Interactions between proteins and genes are often considered essential in the description of biomolecular phenomena and networks of interactions are considered as an entre for a Systems Biology approach. Recently, many works try to extract information by analyzing biomolecular text using natural language processing technology. Previous researches insist that linguistic information is useful to improve the performance in detecting gene interactions. However, previous systems do not show reasonable performance because of low recall. To improve recall without sacrificing precision, this paper proposes a new method for detection of gene interactions based on syntactic relations. Without biomolecular knowledge, our method shows reasonable performance using only small size of training data. Using the format of LLL05(ICML05 Workshop on Learning Language in Logic) data we detect the agent gene and its target gene that interact with each other. In the 1st phase, we detect encapsulation types for each agent and target candidate. In the 2nd phase, we construct verb lists that indicate the interaction information between two genes. In the last phase, to detect which of two genes is an agent or a target, we learn direction information. In the experimental results using LLL05 data, our proposed method showed F-measure of 88% for training data, and 70.4% for test data. This performance significantly outperformed previous methods. We also describe the contribution rate of each phase to the performance, and demonstrate that the first phase contributes to the improvement of recall and the second and last phases contribute to the improvement of precision.