• Title/Summary/Keyword: Hybrid sequencing

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Evidence for VH Gene Replacement in Human Fetal B Cells

  • Lee, Jisoo;Cho, Young Joo;Lipsky, Peter E.
    • IMMUNE NETWORK
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    • v.2 no.2
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    • pp.79-85
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    • 2002
  • Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

Draft genome sequences of Enterococcus faecium JB00008 (KACC 92186P) isolated from Korean fermented soybean paste (Cheonggukjang) (한국 전통유래식품(청국장)에서 분리한 Enterococcus faecium JB00008 (KACC 92186P) 유산균주의 유전체 분석)

  • Park, Jongbin;Jin, Gwi-Deuk;Kim, Eun Bae
    • Korean Journal of Microbiology
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    • v.54 no.2
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    • pp.171-173
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    • 2018
  • Enterococcus faecium was commonly used as a probiotics and feed additives to human and animals because of their beneficial effects. We sequenced the genome of E. faecium JB00008 (KACC 92186P) isolated from a Korean fermented soybean paste (Cheonggukjang) that showed antibacterial activity against Escherichia coli. A 2,847,295-bp draft genome was obtained, and it has in 37.84% G + C content in 34 contigs (length, ${\geq}500bp$).

Development Direction of Reliability-based ROK Amphibious Assault Vehicles (신뢰성 기반 한국군 차기 상륙돌격장갑차 발전방향)

  • Baek, Ilho;Bong, Jusung;Hur, Jangwook
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.20 no.2
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    • pp.14-22
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    • 2021
  • A plan for the development of reliability-based ROK amphibious assault vehicles is proposed. By analyzing the development case of the U.S. EFV, considerations for the successful development of the next-generation Korea Forces amphibious assault vehicle are presented. If the vehicle reliability can be improved to the level of the fourth highest priority electric unit for power units, suspensions, decelerators, and body groups, which have the highest priority among fault frequency items, a system level MTBF of 36.4%↑ can be achieved, and the operational availability can be increased by 3.5%↑. The next-generation amphibious assault vehicles must fulfill certain operating and performance requirements, the underlying systems must be built, and sequencing of the hybrid engine and the modular concept should be considered. Along with big-data- and machine-learning-based failure prediction, machine maintenance based on augmented reality/virtual reality and remote maintenance should be used to improve the ability to maintain combat readiness and reduce lifecycle costs.

Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer (인삼의 유용유전자원 확보를 위한 기능 유전체연구)

  • Yang, Deok-Chun
    • Proceedings of the Ginseng society Conference
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    • 2004.05a
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    • pp.17-28
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    • 2004
  • As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.

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Genomic Analysis of the Xanthoria elegans and Polyketide Synthase Gene Mining Based on the Whole Genome

  • Xiaolong Yuan;Yunqing Li;Ting Luo;Wei Bi;Jiaojun Yu;Yi Wang
    • Mycobiology
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    • v.51 no.1
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    • pp.36-48
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    • 2023
  • Xanthoria elegans is a lichen symbiosis, that inhabits extreme environments and can absorb UV-B. We reported the de novo sequencing and assembly of X. elegans genome. The whole genome was approximately 44.63 Mb, with a GC content of 40.69%. Genome assembly generated 207 scaffolds with an N50 length of 563,100 bp, N90 length of 122,672 bp. The genome comprised 9,581 genes, some encoded enzymes involved in the secondary metabolism such as terpene, polyketides. To further understand the UV-B absorbing and adaptability to extreme environments mechanisms of X. elegans, we searched the secondary metabolites genes and gene-cluster from the genome using genome-mining and bioinformatics analysis. The results revealed that 7 NR-PKSs, 12 HR-PKSs and 2 hybrid PKS-PKSs from X. elegans were isolated, they belong to Type I PKS (T1PKS) according to the domain architecture; phylogenetic analysis and BGCs comparison linked the putative products to two NR-PKSs and three HR-PKSs, the putative products of two NR-PKSs were emodin xanthrone (most likely parietin) and mycophelonic acid, the putative products of three HR-PKSs were soppilines, (+)-asperlin and macrolactone brefeldin A, respectively. 5 PKSs from X. elegans build a correlation between the SMs carbon skeleton and PKS genes based on the domain architecture, phylogenetic and BGC comparison. Although the function of 16 PKSs remains unclear, the findings emphasize that the genes from X. elegans represent an unexploited source of novel polyketide and utilization of lichen gene resources.

Microbial Forensics: Comparison of MLVA Results According to NGS Methods, and Forensic DNA Analysis Using MLVA (미생물법의학: 차세대염기서열분석 방법에 따른 MLVA 결과 비교 및 이를 활용한 DNA 감식)

  • Hyeongseok Yun;Seungho Lee;Seunghyun Lim;Daesang Lee;Sehun Gu;Jungeun Kim;Juhwan Jeong;Seongjoo Kim;Gyeunghaeng Hur;Donghyun Song
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.4
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    • pp.507-515
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    • 2024
  • Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

Identification of relevant differential genes to the divergent development of pectoral muscle in ducks by transcriptomic analysis

  • Fan Li;Zongliang He;Yinglin Lu;Jing Zhou;Heng Cao;Xingyu Zhang;Hongjie Ji;Kunpeng Lv;Debing Yu;Minli Yu
    • Animal Bioscience
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    • v.37 no.8
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    • pp.1345-1354
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    • 2024
  • Objective: The objective of this study was to identify candidate genes that play important roles in skeletal muscle development in ducks. Methods: In this study, we investigated the transcriptional sequencing of embryonic pectoral muscles from two specialized lines: Liancheng white ducks (female) and Cherry valley ducks (male) hybrid Line A (LCA) and Line C (LCC) ducks. In addition, prediction of target genes for the differentially expressed mRNAs was conducted and the enriched gene ontology (GO) terms and Kyoto encyclopedia of genes and genomes signaling pathways were further analyzed. Finally, a protein-to-protein interaction network was analyzed by using the target genes to gain insights into their potential functional association. Results: A total of 1,428 differentially expressed genes (DEGs) with 762 being up-regulated genes and 666 being down-regulated genes in pectoral muscle of LCA and LCC ducks identified by RNA-seq (p<0.05). Meanwhile, 23 GO terms in the down-regulated genes and 75 GO terms in up-regulated genes were significantly enriched (p<0.05). Furthermore, the top 5 most enriched pathways were ECM-receptor interaction, fatty acid degradation, pyruvate degradation, PPAR signaling pathway, and glycolysis/gluconeogenesis. Finally, the candidate genes including integrin b3 (Itgb3), pyruvate kinase M1/2 (Pkm), insulin-like growth factor 1 (Igf1), glucose-6-phosphate isomerase (Gpi), GABA type A receptor-associated protein-like 1 (Gabarapl1), and thyroid hormone receptor beta (Thrb) showed the most expression difference, and then were selected to verification by quantitative real-time polymerase chain reaction (qRT-PCR). The result of qRT-PCR was consistent with that of transcriptome sequencing. Conclusion: This study provided information of molecular mechanisms underlying the developmental differences in skeletal muscles between specialized duck lines.

The Porcine FoxO1, FoxO3a and FoxO4 Genes: Cloning, Mapping, Expression and Association Analysis with Meat Production Traits

  • Yu, Jing;Zhou, Quan-Yong;Zhu, Meng-Jin;Li, Chang-Chun;Liu, Bang;Fan, Bin;Zhao, Shu-Hong
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.627-632
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    • 2007
  • FoxO1, FoxO3a and FoxO4 belong to the FoxO gene family, which play important roles in the PI3K/PKB pathway. In this study, we cloned the porcine FoxO1, FoxO3a and FoxO4 sequences and assigned them to SSC11p11-15, SSC1p13 and SSC xq13 using somatic cell hybrid panel (SCHP) and radiation hybrid panel (IMpRH). RT-PCR results showed that these three genes are expressed in multiple tissues. Sequencing of PCR products from different breeds identified a synonymous T/C polymorphism in exon 2 of FoxO3a. This FoxO3a single nucleotide polymorphism (SNP) can be detected by AvaII restriction enzyme. The allele frequencies of this SNP were investigated in Dahuabai, Meishan, Tongcheng, Yushan, Large White, and Duroc pigs. Association of the genotypes with growth and carcass traits showed that different genotypes of FoxO3a were associated with carcass length and backfat thickness between 6th and 7th ribs (BTR) and drip loss (p<0.05).

Application of Response Surface Methodology in Medium Optimization to Improve Lactic Acid Production by Lactobacillus paracasei SRCM201474 (반응표면분석법을 이용한 Lactobacillus paracasei SRCM201474의 생산배지 최적화)

  • Ha, Gwangsu;Kim, JinWon;Im, Sua;Shin, Su-Jin;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.30 no.6
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    • pp.522-531
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    • 2020
  • The aim of this study was to establish the optimal medium composition for enhancing L(+)-lactic acid (LLA) production using response surface methodology (RSM). Lactobacillus paracasei SRCM201474 was selected as the LLA producer by productivity analysis from nine candidates isolated from kimchi and identified by 16S rRNA gene sequencing. Plackett-Burman design was used to assess the effect of eleven media components on LLA production, including carbon (glucose, sucrose, molasses), nitrogen (yeast extract, peptone, tryptone, beef extract), and mineral (NaCl, K2HPO4, MgSO4, MnSO4) materials. Glucose, sucrose, molasses, and peptone were subsequently chosen as promising media for further optimization studies, and a hybrid design experiment was used to establish their optimal concentrations as glucose 15.48 g/l, sucrose 16.73 g/l, molasses 39.09 g/l, and peptone 34.91 g/l. The coefficient of determination of the equation derived from RSM regression for LLA production was mathematically reliable at 0.9969. At optimum parameters, 33.38 g/l of maximum LLA increased by 193% when compared with MRS broth as unoptimized medium (17.66 g/l). Our statistical model was confirmed by subsequent validation experiments. Increasing the performance of LLA-producing microorganisms and establishing an effective LLA fermentation process can be of particular benefit for bioplastic technologies and industrial applications.

Genetic Species Identification by Sequencing Analysis of Nuclear and Mitochondrial Genes for Albino Misgurnus Species from Korea (우리나라 미꾸리속(genus Misgurnus) 알비노 개체의 미토콘드리아 및 핵 유전자 염기서열 분석에 의한 유전적 동정)

  • Song, Ha-Youn;Moon, Shin-Joo;Kim, Keun-Sik;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.29 no.2
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    • pp.139-145
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    • 2017
  • The spontaneous color mutant, albino individuals of genus Misgurnus, are rarely discovered in Korea and there are difficult to identify morphological species due to lack melanin pigmentation. In this study, we developed a genetic identification method for the species of albino Misgurnus individuals based on phylogenetic analysis by using recombination activating gene 1 (rag1) and cytochrome b (cytb) region of mitochondrial DNA. As a result of molecular phylogenetic analysis, three clades were identified as Misgurnus mizolepis, M. anguillicaudatus and M. mohoity. The homology of the cytb sequences of M. mohoity was best match to that of M. mohoity sequences in GenBank database. As a result of species identification of 25 albino Misgurnus individuals based on the phylogenetic tree, the red-eye type was identified as 16 M. anguillicaudatus and one M. mizolepis. The remaining three individuals were identified as one M. mizolepis ♀${\times}$M. anguillicaudatus ♂, and two M. mohoity ♀${\times}$M. anguillicaudatus ♂, respectively. In addition, the five black-eye type individuals were identified as one M. anguillicaudatus, three M. mizolepis and one M. mohoity. Therefore, this genetic identification method will be an useful techniques for species or hybrid identification in genus Misgurnus.