• Title/Summary/Keyword: Genome analysis

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Expression of an artificial gene encoding a repeated tripeptide lysyl-g1utamyl-tryptophan in Tobacco Plant (담배식물체에서 필수아미노산인 lysyl-glutamyl-tryptophan을 암호화하는 인공유전자의 발현)

  • Lee, Soo-Young;Ra, Kyung-Soo;Baik, Hyung-Suk;Park, Hee-Sung;Cho, Hoon-Sik;Lee, Young-Se;Choi, Jang-Won
    • Journal of Life Science
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    • v.12 no.1
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    • pp.96-105
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    • 2002
  • To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.

Trait Variation and Molecular Characterization of Ds insertional rice lines (벼 Ds 삽입변이 계통의 특성변이 및 분자생물학적 특성)

  • Ahn, Byung-Ohg;Kang, Kyung-Ho;Eun, Moo-Yong;Jeon, Yong-Hee;Yun, Doh-Won;Ji, Hyeon-So;Park, Sung-Han;Nam, Min-Hee;Suh, Seok-Cheol;Lee, Myung-Chul
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.39-47
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    • 2008
  • The 1,874 rice lines were selected from 3,000 Ds insertional mutant pool by Basta herbicide treatment and were surveyed for trait variation and molecular characteristics of genes knocked out by Ds insertion. Compared with "Donjin", an original japonica cultivar used for transformation, Ds insertion mutant pool showed large variation in major agronomic traits including tiller, panicle, and heading etc. Southern blot analysis demonstrated that these lines on the average had two Ds copies in Donjin genome, resulting in 38.4% of one copy, 32.5% of two copies, 16.7% of three copies, and 11.3% of over four copies. GUS analysis showed that 3.9% of lines (73/1,860) had tissue-specific expression in leaves, nodal parts, floral organs such as stigma and pollen, and roots. Data set obtained from agricultural trait variation and molecular characteristics for individual Ds insertional lines would provide researchers with more information for understanding the function of unknown rice genes controlling economically important traits.

Analysis of Genetic Variation Among Popcorn Inbred Lines by SSR Markers (튀김용 옥수수 자식계통들에 대한 유전적 변이성)

  • Jang, Jin-Sun;Chang, Eun-Ha;Sa, Kyu-Jin;Kim, Jong-Hwa;Lee, Ju Kyong
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.405-412
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    • 2011
  • Knowledge of genetic diversity and genetic relationships among inbred lines gives a significant impact on the selection of parental lines for hybrid maize varieties. Genetic diversity and genetic relationships among 86 popcorn inbred lines were analyzed using 50 SSR markers distributed over the whole genome. A total of 256 alleles were identified at all the SSR loci with an average of 5.1 and a range between two and sixteen per locus. The gene diversity values varied from 0.21 to 0.831 with an average of 0.579. The cluster tree generated using the described SSR markers recognized three major groups at 35.8% genetic similarity. Groups I, II, III respectively included 40, 39 and 7 inbred lines. The present study indicates that the SSR markers chosen for this analysis are effective for the assessment of genetic diversity and genetic relationships among 86 popcorn inbred lines in Korea.

MACROD2 Polymorphisms Are Associated with Hypertension in Korean Population (한국인에서의 MACROD2 유전자 다형성과 고혈압 상관성 연구)

  • Ko, Bokyung;Jin, Hyun-Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.57-63
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    • 2019
  • Hypertension is caused by a combination of genetic and environmental factors. In advanced countries, more than 30% of adults have hypertension. Among the genetic factors affecting hypertension, there are reports from European cohort studies that variants of the MACROD2 gene are correlated with blood pressure and the hypertension status. In this study, genetic polymorphisms of the MACROD2 gene region were selected and extracted based on Korean Genome and Epidemiology data, and logistic regression analysis was then performed for the hypertensive state. Linear regression analysis was also performed for the systolic and diastolic blood pressure. As a result, 16 SNPs showed a statistically significant association with a hypertensive state, and 2 SNPs (rs16996211, rs6034240) showed statistical significance, even in blood pressure. The most significant rs16996211 had a relative risk of hypertension of 0.85 (CI: 0.76~0.95, $P=3.1{\times}10^{-3}$), as well as an association with the systolic blood pressure (beta=-0.75, P=0.024) and diastolic blood pressure (beta=-0.59, P=0.01). These results suggest that polymorphisms of the MACROD2 gene are associated with hypertension in both Caucasians and Koreans, and highlight the potential genetic correlations with the pathogenesis of hypertension.

Fine mapping of rice bacterial leaf blight resistance loci on K1 and K2 of Korean races of Xoo (Xanthomonas oryzae) using GWAS analysis

  • Hyeon, Do-Yun;Lee, Jeong-Ro;Jo, Gyu-Taek;Raveendar, Sebastin;Sin, Myeong-Jae;Lee, Gyeong-Jun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.62-62
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    • 2019
  • Bacterial leaf blight(BLB), caused by X. oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice due to its high epidemic potential. Understanding BLB resistance at a genetic level is important to further improve the rice breeding that provides one of the best approaches to control BLB disease. In the present investigation, a collection of 192 accessions was used in the genome-wide association study (GWAS) for BLB resistance loci against four Korean races of Xoo that were represented by the prevailing BLB isolates under Xoo differential system. A total of 192 accessions of rice germplasm were selected on the basis of the bioassay using four isolated races of Xoo such as K1 and K2. The selected accessions was used to prepare 384-plex genotyping by sequencing (GBS) libraries and Illumina HiSeq 2000 pairedend read was used for GBS sequencing. GWAS was conducted using TASSEL 5.0. The TASSEL program uses a mixed linear model (MLM). The results of the bioassay using a selected set of 192 accessions showed that a large number of accessions (93.75%) were resistant to K1 race and K2 resistant germplasm proportion remained between 66.67. The genotypic data produced SNP matrix for a total of 293,379 SNPs. After imputation the missing data was removed, which exhibited 34,724 SNPs for association analysis. GWAS results showed strong signals of association at a threshold of [-log10(P-value)] more than 5 (K1 and K2) for nine of the 39 SNPs, which are plausible candidate loci of resistance genes. These SNP loci were positioned on rice chromosome 2, 9, and 11 for K1 and K2 races. The significant loci detected have also been illustrated and make the CPAS markers for NBS-LRR type disease resistance protein, SNARE domain containing protein, Histone deacetylase 19, NADP-dependent oxidoreductase, and other expressed and unknown proteins. Our results provide a better understanding of the distribution of genetic variation of BLB resistance to Korean pathogen races and breeding of resistant rice.

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Development of Specific SNP Molecular Marker from Thistle in the DNA Sequences of Chloroplast TrnL-F and Matk Region Using HRM Analysis (엉겅퀴의 엽록체 TrnL-F와 Matk 영역 염기서열의 HRM 분석을 통한 특이적 SNP 분자마커의 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Life Science
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    • v.29 no.5
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    • pp.524-529
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    • 2019
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, cosmetics and pharmaceutical industries. However, their phylogenetic origins and names are different from each country and quite often they are mixed each other resulting in the confusion for consumers. Particularly when they are very similar based on their morphological characteristics and distributed, it is extremely difficult to differentiate their origins even by specialists. Therefore, identification of each plant species is important for standardizing herbal medicine. Thistle is a medicinal and perennial plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific thistle species via high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the trnL-F and matK chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different country.

Genome-wide identification and analysis of long noncoding RNAs in longissimus muscle tissue from Kazakh cattle and Xinjiang brown cattle

  • Yan, Xiang-Min;Zhang, Zhe;Liu, Jian-Bo;Li, Na;Yang, Guang-Wei;Luo, Dan;Zhang, Yang;Yuan, Bao;Jiang, Hao;Zhang, Jia-Bao
    • Animal Bioscience
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    • v.34 no.11
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    • pp.1739-1748
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    • 2021
  • Objective: In recent years, long noncoding RNAs (lncRNAs) have been identified in many species, and some of them have been shown to play important roles in muscle development and myogenesis. However, the differences in lncRNAs between Kazakh cattle and Xinjiang brown cattle remain undefined; therefore, we aimed to confirm whether lncRNAs are differentially expressed in the longissimus dorsi between these two types of cattle and whether differentially expressed lncRNAs regulate muscle differentiation. Methods: We used RNA-seq technology to identify lncRNAs in longissimus muscles from these cattle. The expression of lncRNAs were analyzed using StringTie (1.3.1) in terms of the fragments per kilobase of transcript per million mapped reads values of the encoding genes. The differential expression of the transcripts in the two samples were analyzed using the DESeq R software package. The resulting false discovery rate was controlled by the Benjamini and Hochberg's approach. KOBAS software was utilized to measure the expression of different genes in Kyoto encyclopedia of genes and genomes pathways. We randomly selected eight lncRNA genes and validated them by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Results: We found that 182 lncRNA transcripts, including 102 upregulated and 80 downregulated transcripts, were differentially expressed between Kazakh cattle and Xinjiang brown cattle. The results of RT-qPCR were consistent with the sequencing results. Enrichment analysis and functional annotation of the target genes revealed that the differentially expressed lncRNAs were associated with the mitogen-activated protein kinase, Ras, and phosphatidylinositol 3-kinase (PI3k)/Akt signaling pathways. We also constructed a lncRNA/mRNA coexpression network for the PI3k/Akt signaling pathway. Conclusion: Our study provides insights into cattle muscle-associated lncRNAs and will contribute to a more thorough understanding of the molecular mechanism underlying muscle growth and development in cattle.

Molecular Analysis of the Interaction between Human PTPN21 and the Oncoprotein E7 from Human Papillomavirus Genotype 18

  • Lee, Hye Seon;Kim, Min Wook;Jin, Kyeong Sik;Shin, Ho-Chul;Kim, Won Kon;Lee, Sang Chul;Kim, Seung Jun;Lee, Eun-Woo;Ku, Bonsu
    • Molecules and Cells
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    • v.44 no.1
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    • pp.26-37
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    • 2021
  • Human papillomaviruses (HPVs) cause cellular hyperproliferation-associated abnormalities including cervical cancer. The HPV genome encodes two major viral oncoproteins, E6 and E7, which recruit various host proteins by direct interaction for proteasomal degradation. Recently, we reported the structure of HPV18 E7 conserved region 3 (CR3) bound to the protein tyrosine phosphatase (PTP) domain of PTPN14, a well-defined tumor suppressor, and found that this intermolecular interaction plays a key role in E7-driven transformation and tumorigenesis. In this study, we carried out a molecular analysis of the interaction between CR3 of HPV18 E7 and the PTP domain of PTPN21, a PTP protein that shares high sequence homology with PTPN14 but is putatively oncogenic rather than tumor-suppressive. Through the combined use of biochemical tools, we verified that HPV18 E7 and PTPN21 form a 2:2 complex, with a dissociation constant of 5 nM and a nearly identical binding manner with the HPV18 E7 and PTPN14 complex. Nevertheless, despite the structural similarities, the biological consequences of the E7 interaction were found to differ between the two PTP proteins. Unlike PTPN14, PTPN21 did not appear to be subjected to proteasomal degradation in HPV18-positive HeLa cervical cancer cells. Moreover, knockdown of PTPN21 led to retardation of the migration/invasion of HeLa cells and HPV18 E7-expressing HaCaT keratinocytes, which reflects its protumor activity. In conclusion, the associations of the viral oncoprotein E7 with PTPN14 and PTPN21 are similar at the molecular level but play different physiological roles.

Identification of a Single Nucleotide Polymorphism (SNP) Marker for the Detection of Enhanced Honey Production in Hoenybee (수밀력 우수 꿀벌 계통 판별을 위한 계통 특이 분자마커 개발)

  • Kim, Hye-Kyung;Lee, Myeong-Lyeol;Lee, Man-Young;Choi, Yong-Soo;Kim, Dongwon;Kang, Ah Rang
    • Journal of Apiculture
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    • v.32 no.3
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    • pp.147-154
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    • 2017
  • Honeybees (Apis mellifera) are common pollinators and important insects studied in agriculture, ecology and basic research. Recently, RDA (Rural Development Administration) and YIRI (Yecheon-gun Industrial Insect Research Institute) have been breeding a triple crossbred honey bee named Jangwon, which have the ability to produce superior quality honey. In this study, we identified a single nucleotide polymorphism (SNP) marker in the genome of Jangwon honeybee, particularly, in the paternal line (D line). Initially, we performed Sequence-Based Genotyping (SBG) using the Illumina Hiseq 2500 in 5 honeybee inbred lines; A, C, D, E, and F; and obtained 1,029 SNPs. Seventeen SNPs for each inbred line were generated and selected after further filtering of the SNP dataset. The 17 SNP markers validated by performing TaqMan probe-based real-time PCR and genotyping analysis was conducted. Genotyping analysis of the 5 honeybee inbred lines and one hybrid line, $D{\times}F$, revealed that one set of SNP marker, AmD9, precisely discriminated the inbred line D from the others. Our results suggest that the identified SNP marker, AmD9, is successful in distinguishing the inbred honeybee lines D, and can be directly used for genotyping and breeding applications.

First Report of Tomato Spotted Wilt Virus in Angelica acutiloba (당귀에서 발생한 토마토반점위조바이러스의 감염 첫 보고)

  • Kwak, Hae-Ryun;Hong, Su-Bin;Choi, Hyeon-Yong;Park, Gosoo;Hur, On-Sook;Byun, Hee-Seong;Choi, Hong-Soo;Kim, Mikyeong
    • Research in Plant Disease
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    • v.27 no.2
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    • pp.84-90
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    • 2021
  • In June 2019, Angelica acutiloba plants showing virus-like symptoms such as chlorotic local lesion and mosaic on the leaves were found in a greenhouse in Nonsan, South Korea. To identify the causal virus, we collected 6 symptomatic A. acutiloba leaf samples and performed reverse transcription polymerase chain reaction (RT-PCR) analysis using specific detection primers for three reported viruses including tomato spotted wilt virus (TSWV). RT-PCR results showed that five symptomatic samples were positive for TSWV. Mechanical sap inoculation of one of the collected TSWV isolate (TSWV-NS-AG28) induced yellowing, chlorosis and mosaic symptoms in A. acutiloba and necrotic local lesions and mosaic in Solanaceae species. Phylogenetic analysis based on the complete genome sequences showed that TSWV-NS-AG28 had a maximum nucleotide identity with TSWVNS-BB20 isolated from butterbur in Nonsan, South Korea. To our knowledge, this is the first report of TSWV infection in A. acutiloba.