• Title/Summary/Keyword: Whole-genome sequence

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Complete Genome Sequence and Antimicrobial Activities of Bacillus velezensis MV2 Isolated from a Malva verticillate Leaf (아욱 잎에서 분리한 Bacillus velezensis MV2의 유전체 염기서열 분석과 항균활성능 연구)

  • Lee, Hyeonju;Jo, Eunhye;Kim, Jihye;Moon, Keumok;Kim, Min Ji;Shin, Jae-Ho;Cha, Jaeho
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.111-119
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    • 2021
  • A bacterial strain isolated from a Malva verticillata leaf was identified as Bacillus velezensis MV2 based on the 16S rRNA sequencing results. Complete genome sequencing revealed that B. velezensis MV2 possessed a single 4,191,702-bp contig with 45.57% GC content. Generally, Bacillus spp. are known to produce diverse antimicrobial compounds including bacteriocins, polyketides, and non-ribosomal peptides. Antimicrobial compounds in the B. velezensis MV2 were extracted from culture supernatants using hydrophobic interaction chromatography. The crude extracts showed antimicrobial activity against both gram-positive bacteria and gram-negative bacteria; however, they were more effective against gram-positive bacteria. The extracts also showed antifungal activity against phytopathogenic fungi such as Fusarium fujikuroi and F. graminearum. In time-kill assays, these antimicrobial compounds showed bactericidal activity against Bacillus cereus, used as indicator strain. To predict the type of antimicrobial compounds produced by this strain, we used the antiSMASH algorithm. Forty-seven secondary metabolites were predicted to be synthesized in MV2, and among them, fourteen were identified with a similarity of 80% or more with those previously identified. Based on the antimicrobial properties, the antimicrobial compounds may be nonribosomal peptides or polyketides. These compounds possess the potential to be used as biopesticides in the food and agricultural industry as an alternative to antibiotics.

Characterization of the cloned RNA1 gene of Saccharomyces cerevisiae (Cloning된 효모의 RNAI 유전자의 특성에 관하여)

  • Song, Young-Hwan;Kim, Dae-Young;Kim, Jin-Kyung
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.93-101
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    • 1993
  • The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

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Development of SSLP Marker Targeted to P34 Null Gene in Soybean (콩 P34 단백질 결핍 유전자를 이용한 SSLP 마커 개발)

  • Yang, Kiwoung;Ko, Jong-Min;Lee, Young-Hoon;Jeon, Myeong Gi;Jung, Chan-Sik;Baek, In-Youl;Kim, Hyun-Tae;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.502-506
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    • 2010
  • Soybean seed possesses about 15 allergenic proteins recognized by IgEs from soy-sensitive human. The allergenic impact of soybean proteins limit its extensive usage in a broad range of processed foods. Soybean protein P34 or Gly m Bd 30k of the cysteine protease family is one of the major allergen of the soybean seed. P34-null soybean, PI567476, was identified among soybean (Glycine max & Glycine soja Sieb. and Zucc) of approximately 16,226 accessions from USDA soybean germplasm screened. Also, for P34 gene (Williams 82; whole genome sequence cultivar) and P34 null gene (PI567476) comparative analysis of sequences listed in the NCBI database showed the presence of a SSLP (Simple Sequence Length Polymorphism) of 4 base pair. So, a SSLP marker was designed to reveal the polymorphism of the locus. In this study, a population of 339 $F_2$ recombinant inbred lines generated by cross between Taekwang (Glycine max) and PI567476 was used to select $F_{2:3}$ plant of a P34 null gene. The result separation rate Taekwang type, heterozygous type and PI567476 type were shown in 85: 187: 67 since single gene is concerned in as the separation rate of 1:2:1 in $X^2{_{0.05}}=5.99$, df=2. In future, selected plant will identify protein level, whether P34 null protein is equal to P34 null gene.

Current Status of Cattle Genome Sequencing and Analysis using Next Generation Sequencing (차세대유전체해독 기법을 이용한 소 유전체 해독 연구현황)

  • Choi, Jung-Woo;Chai, Han-Ha;Yu, Dayeong;Lee, Kyung-Tai;Cho, Yong-Min;Lim, Dajeong
    • Journal of Life Science
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    • v.25 no.3
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    • pp.349-356
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    • 2015
  • Thanks to recent advances in next-generation sequencing (NGS) technology, diverse livestock species have been dissected at the genome-wide sequence level. As for cattle, there are currently four Korean indigenous breeds registered with the Domestic Animal Diversity Information System of the Food and Agricultural Organization of the United Nations: Hanwoo, Chikso, Heugu, and Jeju Heugu. These native genetic resources were recently whole-genome resequenced using various NGS technologies, providing enormous single nucleotide polymorphism information across the genomes. The NGS application further provided biological such that Korean native cattle are genetically distant from some cattle breeds of European origins. In addition, the NGS technology was successfully applied to detect structural variations, particularly copy number variations that were usually difficult to identify at the genome-wide level with reasonable accuracy. Despite the success, those recent studies also showed an inherent limitation in sequencing only a representative individual of each breed. To elucidate the biological implications of the sequenced data, further confirmatory studies should be followed by sequencing or validating the population of each breed. Because NGS sequencing prices have consistently dropped, various population genomic theories can now be applied to the sequencing data obtained from the population of each breed of interest. There are still few such population studies available for the Korean native cattle breeds, but this situation will soon be improved with the recent initiative for NGS sequencing of diverse native livestock resources, including the Korean native cattle breeds.

Chromosomal Localization and Mutation Detection of the Porcine APM1 Gene Encoding Adiponectin (Adiponectin을 암호화하는 돼지 APM1 유전자의 염색체상 위치파악과 돌연변이 탐색)

  • Park, E.W.;Kim, J.H.;Seo, B.Y.;Jung, K.C.;Yu, S.L.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.537-546
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    • 2004
  • Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.

Identification of a Second Type of AHL-Lactonase from Rhodococcus sp. BH4, belonging to the α/β Hydrolase Superfamily

  • Ryu, Du-Hwan;Lee, Sang-Won;Mikolaityte, Viktorija;Kim, Yea-Won;Jeong, Haeyoung;Lee, Sang Jun;Lee, Chung-Hak;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.937-945
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    • 2020
  • N-acyl-homoserine lactone (AHL)-mediated quorum sensing (QS) plays a major role in development of biofilms, which contribute to rise in infections and biofouling in water-related industries. Interference in QS, called quorum quenching (QQ), has recieved a lot of attention in recent years. Rhodococcus spp. are known to have prominent quorum quenching activity and in previous reports it was suggested that this genus possesses multiple QQ enzymes, but only one gene, qsdA, which encodes an AHL-lactonase belonging to phosphotriesterase family, has been identified. Therefore, we conducted a whole genome sequencing and analysis of Rhodococcus sp. BH4 isolated from a wastewater treatment plant. The sequencing revealed another gene encoding a QQ enzyme (named jydB) that exhibited a high AHL degrading activity. This QQ enzyme had a 46% amino acid sequence similarity with the AHL-lactonase (AidH) of Ochrobactrum sp. T63. HPLC analysis and AHL restoration experiments by acidification revealed that the jydB gene encodes an AHL-lactonase which shares the known characteristics of the α/β hydrolase family. Purified recombinant JydB demonstrated a high hydrolytic activity against various AHLs. Kinetic analysis of JydB revealed a high catalytic efficiency (kcat/KM) against C4-HSL and 3-oxo-C6 HSL, ranging from 1.88 x 106 to 1.45 x 106 M-1 s-1, with distinctly low KM values (0.16-0.24 mM). This study affirms that the AHL degrading activity and biofilm inhibition ability of Rhodococcus sp. BH4 may be due to the presence of multiple quorum quenching enzymes, including two types of AHL-lactonases, in addition to AHL-acylase and oxidoreductase, for which the genes have yet to be described.

Development and Application of DNA Analysis Method for Identificaion of Main Ingredients in Starch (전분의 주원료 판별을 위한 유전자 분석법 개발 및 적용)

  • Park, Yong-Chjun;Kim, Mi-Ra;Kim, Yong-Sang;Lee, Ho-Yeon;Kim, Kyu-Heon;Lee, Jae-Hwang;Kim, Jae-I;Lee, Sang-Jae;Lee, Hwa-Jung
    • Journal of Food Hygiene and Safety
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    • v.28 no.2
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    • pp.181-187
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    • 2013
  • Identification of main ingredients in starches has been investigated using physicochemical analysis method mainly. However, physicochemical properties such as particle size have limitations in determining the differences among mixed starches. Therefore, we developed a molecular biological method to identify materials used in starch, as a sample, 11 kinds of starches including sweet potato starch, potato starch, corn starch, and tapioca starch. DNeasy plant mini kit, magnetic DNA purification system, and CTAB methods were used to extract DNA from samples. After gene extraction, whole genome amplification (WGA) was performed to amplify the extracted DNA. Species-specific primers were used as followings: ib-286-F/ib-286-R (105 bp), Pss 01n-5'/Pss 01n-3' (216 bp), SS11b 3-5'/SS11b 3-3' (114 bp), and SSRY26-F/SSRY26-R (121 bp) gene for sweet potato, potato, corn, and tapioca, respectively. In this study, we could confirm the main ingredients using WGA and PCR method.

Strategies to Increase Domestic Lettuce Circulations through Improving Valuable End-User Traits (고부가가치 맞춤형 상추품종 개발을 통한 국내 상추유통 제고 전략)

  • Kim, Tae-Sung;Jang, Young-Hee;Hwang, Hee-Joong
    • Journal of Distribution Science
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    • v.16 no.8
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    • pp.63-68
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    • 2018
  • Purpose - Lettuce (Lactuca sativ L.) is one of the economically important vegetable crops, which worldwide market value is over 100 billion U.S. dollar. In Korea, about 89.7 kilo ton of lettuce was produced in 3400ha in 2016, recoded as No. 1 vegetable crop in domestic green house production. However, recently, domestic lettuce production and cultivation areas are all getting decreased. Thus, novel approaches are needed to be implemented to revive the production. Research design, data and methodology - In this review paper, we first prioritized the end-user traits which are imperative to positively stimulate the domestic lettuce market and discussed relevant genomics strategies. Especially, we assessed a possibility whether school meal program would be a potential niche market. Results - The genomics technologies, which become widely applied in the crop biotechnology since 2008 when next generation sequencing method was developed, may be a good solution in the crop improvement, efficiently gathering valuable information of agriculturally useful traits. Significantly, in lettuce, the high quality whole genome sequence, based on Lactuca sativa cv. Salinas, is publically available and this genomics platform, thus, would be implemented in lettuce breeding program to innovate relevant end-user traits both for the farmers and customers, including the disease resistance to the Fusarium wilt, productivity under hot weather conditions, various nutritional qualities and so forth. These improvements will boost domestic lettuce industries in the near future. Conclusions - Due to the nutritional distinctions comparing to the western style lettuces, domestic leaf lettuces could be one of the important vegetables in the school meal programs. To make it happen, we would better devise diverse recipes to make a salad with it, instead of only using as a wrap vegetable. Meanwhile, novel lettuce varieties need to be developed, which are favorable to the students and also easy to be handled with while processing. Overall, to achieve international competence in the lettuce industries, we need to create elite lettuce varieties that satisfies domestic farmers as well as customers, suitable to various niche markets, such as school meal program. Thus, efficient breeding programs using genomics approaches should be established in advance and careful monitoring on the preference of the related customers for a niche market be continued persistently.

First Report of Tomato Spotted Wilt Virus on Iris domestica in South Korea (약용작물 범부채에 발생한 Tomato Spotted Wilt Virus 국내 첫 보고)

  • Chung, Bong Nam;Yoon, Ju-Yeon;Cho, In-Sook
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.32-37
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    • 2021
  • In May 2020, necrosis and necrotic ring patterns were observed on leaves of three of 140 Iris domestica plants in a demonstration garden in Wanju, Jeollabuk-do. Three symptomatic plants were found to be infected by tomato spotted wilt virus (TSWV). To analyze the whole genomic sequence of one TSWV isolate, 'Blackberry lily-kr1', L, M, and S genome segments were sequenced and analyzed by comparison of nucleotide sequences of the three segments with corresponding sequences of other TSWV isolates. 'Blackberry lily-kr1' isolate was most closely related to 'JJ' isolate (MF159046) or 'HJ' isolate (LC273305) in the L segment, and to 'JJ' isolate (MF159058 and KY021439) in the M and S segments, respectively. Phylogenetic analysis by Maximum likelihood method using MEGA X program with 'Blackberry lily-kr1' isolate showed high relationship with 'JJ' pepper isolate or 'HJ' Humulus japonicas isolate in the all three segment. Necrosis and double ring patterns on leaves were formed in the glasshouse after inoculation of healthy I. domestica plants with sap of 'Blackberry lily-kr1'-infected Nicotiana rustica plants. This result suggests that I. domestica plants showing necrotic ring patterns in the open field are caused by TSWV infection. This is the first report of TSWV infection of I. domestica in Korea.

Design and Implementation of Memory-Centric Computing System for Big Data Analysis

  • Jung, Byung-Kwon
    • Journal of the Korea Society of Computer and Information
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    • v.27 no.7
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    • pp.1-7
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    • 2022
  • Recently, as the use of applications such as big data programs and machine learning programs that are driven while generating large amounts of data in the program itself becomes common, the existing main memory alone lacks memory, making it difficult to execute the program quickly. In particular, the need to derive results more quickly has emerged in a situation where it is necessary to analyze whether the entire sequence is genetically altered due to the outbreak of the coronavirus. As a result of measuring performance by applying large-capacity data to a computing system equipped with a self-developed memory pool MOCA host adapter instead of processing large-capacity data from an existing SSD, performance improved by 16% compared to the existing SSD system. In addition, in various other benchmark tests, IO performance was 92.8%, 80.6%, and 32.8% faster than SSD in computing systems equipped with memory pool MOCA host adapters such as SortSampleBam, ApplyBQSR, and GatherBamFiles by task of workflow. When analyzing large amounts of data, such as electrical dielectric pipeline analysis, it is judged that the measurement delay occurring at runtime can be reduced in the computing system equipped with the memory pool MOCA host adapter developed in this research.