• Title/Summary/Keyword: Genome Analysis

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Nucleus-Selective Expression of Laccase Genes in the Dikaryotic Strain of Lentinula edodes

  • Ha, Byeongsuk;Lee, Sieun;Kim, Sinil;Kim, Minseek;Moon, Yoon Jung;Song, Yelin;Ro, Hyeon-Su
    • Mycobiology
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    • v.45 no.4
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    • pp.379-384
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    • 2017
  • In mating of Lentinula edodes, dikaryotic strains generated from certain monokaryotic strains such as the B2 used in this study tend to show better quality of fruiting bodies regardless of the mated monokaryotic strains. Unlike B2, dikaryotic strains generated from B16 generally show low yields, with deformed or underdeveloped fruiting bodies. This indicates that the two nuclei in the cytoplasm do not contribute equally to the physiology of dikaryotic L. edodes, suggesting an expression bias in the allelic genes of the two nuclei. To understand the role of each nucleus in dikaryotic strains, we investigated single nucleotide polymorphisms (SNPs) in laccase genes of monokaryotic strains to reveal nuclear origin of the expressed mRNAs in dikaryotic strain. We performed reverse transcription PCR (RT-PCR) analysis using total RNAs extracted from dikaryotic strains (A5B2, A18B2, and A2B16) as well as from compatible monokaryotic strains (A5, A18, and B2 for A5B2 and A18B2; A2 and B16 for A2B16). RT-PCR results revealed that Lcc1, Lcc2, Lcc4, Lcc7, and Lcc10 were the mainly expressed laccase genes in the L. edodes genome. To determine the nuclear origin of these laccase genes, the genomic DNA sequences in monokaryotic strains were analyzed, thereby revealing five SNPs in Lcc4 and two in Lcc7. Subsequent sequence analysis of laccase mRNAs expressed in dikaryotic strains revealed that these were almost exclusively expressed from B2-originated nuclei in A5B2 and A18B2 whereas B16 nucleus did not contribute to laccase expression in A2B16 strain. This suggests that B2 nucleus dominates the expression of allelic genes, thereby governing the physiology of dikaryons.

Characterization of the Neurospora crassa rcm-1 Mutants (Neurospora crassa rcm-1 돌연변이체의 특성)

  • Kim Sang-Rae;Lee Bheong-Uk
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.246-254
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    • 2005
  • Analysis of the complete genome of Neurospora crassa reveals that at least 19 proteins contain tetratricopeptide repeat (TPR) motifs. One of them shows over $60\%$ homology to Ssn6 of Saccharomyces cerevisiae, a universal repressor that mediates repression of genes involved in various cellular processes. Mutant strains generated by RIP (repeat-induced point mutation) process showed four distinctive vegetative growth patterns and slow growth in various rates. Firstly, a mutant showed denser mycelial growth, yellow, csp, and looked like ropy mutant. Secondly, slower growth, dense mycelial, and conidial phenotype. Thirdly, extremely slower growth and aconidial. And finally, flat, tittle aerial hyphae, acon, and similar with a rco-1 RIP mutant. They are all male-fertile, yet female-sterile and produced little or no perithecium. It seems that various phenotypes were occurred depending upon mostly likely, the degree of RIP. These results indicate that this gene may be involved in several cellular possess during vegetative growth, and asexual and sexual development. Therefore it is pleiotropic. Sequence analysis of cDNA shows that it encodes a putative 102 kDa protein composed of 917 amino acids, and has six introns. It is designated rcm-1 (regulation of conidiation and morphology).

Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.147-152
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    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

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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.