• 제목/요약/키워드: Chromosomal technology

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Dynamics of Viral and Host 3D Genome Structure upon Infection

  • Meyer J. Friedman;Haram Lee;Young-Chan Kwon;Soohwan Oh
    • Journal of Microbiology and Biotechnology
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    • 제32권12호
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    • pp.1515-1526
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    • 2022
  • Eukaryotic chromatin is highly organized in the 3D nuclear space and dynamically regulated in response to environmental stimuli. This genomic organization is arranged in a hierarchical fashion to support various cellular functions, including transcriptional regulation of gene expression. Like other host cellular mechanisms, viral pathogens utilize and modulate host chromatin architecture and its regulatory machinery to control features of their life cycle, such as lytic versus latent status. Combined with previous research focusing on individual loci, recent global genomic studies employing conformational assays coupled with high-throughput sequencing technology have informed models for host and, in some cases, viral 3D chromosomal structure re-organization during infection and the contribution of these alterations to virus-mediated diseases. Here, we review recent discoveries and progress in host and viral chromatin structural dynamics during infection, focusing on a subset of DNA (human herpesviruses and HPV) as well as RNA (HIV, influenza virus and SARS-CoV-2) viruses. An understanding of how host and viral genomic structure affect gene expression in both contexts and ultimately viral pathogenesis can facilitate the development of novel therapeutic strategies.

Current status of whole-genome sequences of Korean angiosperms

  • Jongsun PARK;Yunho YUN;Hong XI;Woochan KWON;Janghyuk SON
    • 식물분류학회지
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    • 제53권3호
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    • pp.181-200
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    • 2023
  • Owing to the rapid development of sequencing technologies, more than 1,000 plant genomes have been sequenced and released. Among them, 69 Korean plant taxa (85 genome sequences) contain at least one whole-genome sequence despite the fact that some samples were not collected in Korea. The sequencing-by-synthesis method (next-generation sequencing) and the PacBio (third-generation sequencing) method were the most commonly used in studies appearing in 65 publications. Several scaffolding methods, such as the Hi-C and 10x types, have also been used for pseudo-chromosomal assembly. The most abundant families among the 69 taxa are Rosaceae (10 taxa), Brassicaceae (7 taxa), Fabaceae (7 taxa), and Poaceae (7 taxa). Due to the rapid release of plant genomes, it is necessary to assemble the current understanding of Korean plant species not only to understand their whole genomes as our own plant resources but also to establish new tools for utilizing plant resources efficiently with various analysis pipelines, including AI-based engines.

한우의 유전체 표지인자 활용 개체 혈연관계 추정 (Prediction of Genomic Relationship Matrices using Single Nucleotide Polymorphisms in Hanwoo)

  • 이득환;조충일;김내수
    • Journal of Animal Science and Technology
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    • 제52권5호
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    • pp.357-366
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    • 2010
  • 한우의 유전체 전장의 정보를 Illumina BeadArray$^{TM}$ Bovine SNP50 assay를 이용하여 단일염기다형 현상을 조사한 결과, 유전적 다양성을 보이는 좌위가 약 32,567 좌위 이상에서 다양성을 보이고 있었으며 약 5,554 좌위에서 다양성이 조사되지 않았다. 이는 조사된 자료의 가계집단의 수가 크게 제한되었기 때문에 기인될 수 있으며 또 다른 원인으로는 한우 종축집단의 크기가 작을 수 있다는 현상을 반증한다고 사료된다. 유전분석의 기초가 되는 혈통기록에 의한 개체간 혈연관계를 유전체 정보에 의한 혈연관계와 비교하여 본 결과, 유전체 정보에 의한 혈연관계의 크기가 혈통기록에 의한 혈연관계보다 좀 더 정확하게 추정될 수 있다는 장점이 있으며 혈통기록상의 오류로 그릇된 혈연관계의 크기를 유전체 정보를 통하여 보완할 수 있다는 장점이 있다. 이러한 장점을 활용하면 유전체정보를 이용한 유전능력 평가의 정확성을 크게 향상시킬 수 있을 것으로 사료되었다.

Avirulence Gene Diversity of Xanthomonas axonopodis pv. glycines Isolated in Korea

  • Park, Hyoung-Joon;Han, Sang-Wook;Oh, Chang-Sik;Lee, Seung-Don;Ra, Dong-Soo;Lee, Suk-Ha;Heu, Sung-Gi
    • Journal of Microbiology and Biotechnology
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    • 제18권9호
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    • pp.1500-1509
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    • 2008
  • The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.

Genome wide association study of fatty acid composition in Duroc swine

  • Viterbo, Vanessa S.;Lopez, Bryan Irvine M.;Kang, Hyunsung;Kim, Hoonseop;Song, Choul-won;Seo, Kang Seok
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권8호
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    • pp.1127-1133
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    • 2018
  • Objective: Genome wide association study was conducted to identify and validate candidate genes associated with fatty acid composition of pork. Methods: A total of 480 purebreed Duroc pigs were genotyped using IlluminaPorcine60k bead chips while the association test was implemented following genome-wide rapid association using Mixed Model and Regression-Genomic Control (GRAMMAR-GC) approach. Results: A total of 25, 29, and 16 single nucleotide polymorphisms (SNPs) were significantly associated with stearic (18:0), oleic (18:1) and saturated fatty acids (SFA), respectively. Genome wide significant variants were located on the same region of swine chromosome 14 (SSC14) that spanned from 120 to 124 Mb. Top SNP ALGA008191 was located at 5 kb near the stearoyl-CoA desaturase (SCD) gene. This gene is directly involved in desaturation of stearic acid into oleic acid. General relationship of significant SNPs showed high linkage disequilibrium thus genome-wide signals was attributed to SCD gene. However, understanding the role of other genes like elongation of very long chain fatty acids-3 (ELOVL3) located on this chromosomal segment might help in further understanding of metabolism and biosynthesis of fatty acids. Conclusion: Overall, this study provides evidence that validates SCD gene as strong candidate gene associated with fatty acid composition in Duroc pigs. Moreover, this study confirms significant SNPs near ELOVL3 gene.

Cloning of the Endoglucanase Gene from Actinomyces sp. 40 in Escherichia coli and Some Properties of the Gene Products

  • Min, Hae-Ki;Choi, Yun-Jaie;Cho, Kwang-Keun;Ha, Jong-Kyu;Woo, Jung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제4권2호
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    • pp.102-107
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    • 1994
  • The $\beta$-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5$\alpha$ with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3AI and ligated into pUC19 for the transformation of Escherichia coli DH5$\alpha$. Positive clones of $\beta$-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3AI insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5$\alpha$ (pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0$\sim$5.0 and 55$^{\circ}C$, respectively. The cloned enzyme was stable at 55$^{\circ}C$ or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).

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Direct Analysis of the Transcription of Escherichia coli rnpB Gene Harbored in a Multicopy Plasmid during Bacterial Growth

  • Park, Jeong-Won;Jung, Young-Hwan;Park, Bo-Hyun;Jeoung, Yeon-Hee;Lee, Young-Hoon
    • BMB Reports
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    • 제29권3호
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    • pp.221-224
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    • 1996
  • To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

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Screening and Characterization of Secretion Signals from Lactococcus lactis ssp. cremoris LM0230

  • Jeong, Do-Won;Choi, Youn-Chul;Lee, Jung-Min;Seo, Jung-Min;Kim, Jeong-Hwan;Lee, Jong-Hoon;Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • 제14권5호
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    • pp.1052-1056
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    • 2004
  • A secretion signal sequence-selection vector (pGS40) was constructed based on an $\alpha$-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore $\alpha$-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgamo (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.

Sequence Characterization, Expression Profile, Chromosomal Localization and Polymorphism of the Porcine SMPX Gene

  • Guan, H.P.;Fan, B.;Li, K.;Zhu, M.J.;Yerle, M.;Liu, Bang
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권7호
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    • pp.931-937
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    • 2006
  • The full-length cDNA of the porcine SMPX gene was obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with both human and mouse. The promoter of SMPX was sequenced and then analyzed to find the promoter binding sites. The reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that SMPX has a high level of expression in heart and skeletal muscle, a very low expression in lung and spleen and no expression in liver, kidney, fat and brain. Moreover, SMPX has a differential expression level in skeletal muscle, the expression in 65-day embryos being higher than other stages. The porcine SMPX was mapped to SSCXp24 by using a somatic cell hybrid panel (SCHP) and was found closely linked to SW1903 using the radiation hybrid panel IMpRH. An A/G single nucleotide polymorphism (PCR-RFLP) in the 3'-untranslated region (3'-UTR) was detected in eight breeds. The analysis of allele frequency distribution showed that introduced pig breeds (Duroc and Large White) have a higher frequency of allele A while in the Chinese indigenous pig breeds (Qingping pig, Lantang pig, YushanBlack pig, Large Black-White pig, Small Meishan) have a higher frequencies of allele G. The association analysis using an experimental population (188 pigs), which included two cross-bred groups and three pure-blood groups, suggested that the SNP genotype was associated with intramuscular fat content.

The New LM-PCR/Shifter Method for the Genotyping of Microorganisms Based on the Use of a Class IIS Restriction Enzyme and Ligation-Mediated PCR

  • Krawczyk, Beata;Leibner-Ciszak, Justyna;Stojowska, Karolina;Kur, Jozef
    • Journal of Microbiology and Biotechnology
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    • 제21권12호
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    • pp.1336-1344
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    • 2011
  • This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.