• Title/Summary/Keyword: cDNA sequence

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Production and Characterization of Mannanase from a Bacillus sp. YB-1401 Isolated from Fermented Soybean Paste (된장 분리균 Bacillus sp. YB-1401의 Mannanase 생산성과 효소특성)

  • Joen, Ho Jin;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.99-105
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    • 2014
  • A Bacillus strain capable of hydrolyzing locust bean gum was isolated as a producer of extracellular mannanase by way of an enrichment culture in an acidic medium from homemade soybean pastes. The isolate YB-1401 showed a biochemical identity of 61.1% with Brevibacillus laterosporus, while the nucleotide sequence of its 16S rDNA had the highest similarity with that of Bacillus amyloliquefaciens. The mannanase productivity of the Bacillus sp. YB-1401 was drastically increased by mannans. Particularly, maximum mannanase productivity was reached at approximately 265 U/ml in LB medium supplemented with konjac glucomannan (4.0%). The mannanase was the most active at $55^{\circ}C$ and pH 5.5. Mannanase activity was completely maintained after pre-incubation at pH 3.5 to 11.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannobiose and mannotriose for LBG, guar gum or mannooligosaccharides. A small amount of mannose was also detected in the hydrolyzates.

Additional mitochondrial DNA sequences from the dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), an endangered species in South Korea

  • Hwang, Eun Ju;Jeong, Su Yeon;Wang, Ah Rha;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.2
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    • pp.31-41
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    • 2018
  • The dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), is an endangered insect in South Korea. Previously, partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of this species in South Korea. In this study, we additionally collected C. tripartitus (n = 35) from one previous locality and two new localities, sequenced COI and CytB genes, and combined these with preexisting data for population genetic analysis. Sequence divergence of current samples showed slightly lower values [4.86% (32 bp) for COI and 4.16% (18 bp) for CytB] than that in the previous study. Nucleotide diversity (${\pi}$) ranged from 0.005336 (Gulupdo) to 0.020756 (Seogwi-dong) in COI and 0.009060 (Aewol-eup) to 0.017464 (Seogwi-dong) in CytB. Seogwi-dong samples that showed the highest ${\pi}$ in the previous study also showed the highest ${\pi}$ in this study for both gene sequences. The newly investigated Gulupdo samples had the lowest haplotype diversity for both gene sequences. They also had the lowest ${\pi}$ for COI and the second lowest ${\pi}$ for CytB. On the other hand, the newly added Haean-dong sample had relatively higher diversity estimates. Gene flow among populations was high, although significant difference was only detected between Gulupdo and Anmado or between Gulupdo and Seogwi-dong for COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one major issue regarding conservation seems not to be recovery of genetic diversity.

Pathgenicity on Ginseng and Sequence Assays of Ilyonectria radicicola Isolated from Chestnut Rhizosphere Soils (밤나무 근권토양에서 분리한 Ilyonectria radicicola 균주의 인삼에 대한 병원성 및 유전적 분석)

  • Seo, Mun Won;Song, Jeong Young;Kim, Sun Ick;Oh, Sang Keun;Kim, Hong Gi
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.4
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    • pp.302-307
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    • 2018
  • Background: A soil-borne pathogenic fungus, Ilyonectria radicicola (Cylindrocarpon destructans) causes root rot on ginseng (Panax ginseng C. A. Meyer) and is known to attack many other plants. The Nectria/Neonectria radicicola complex has been renamed as the I. radicicola complex after analysis of its multi-gene relatedness and morphological characteristics. The fungi in this complex have been reclassified into 16 species under the genus Ilyonectria based on characteristics analysis Methods and Results: To obtain useful data from the Korean ginseng root rot, I. radicicola was isolated from the rhizosphere soils of the chestnut tree. They were identified through a pathogenicity test and a survey of the morphological features. The existence of I. radicicola in soil samples was confirmed by PCR detections using nested PCR with species-specific primer sets. These were subsequenctly isolated on semi-selective media from PCR-positive soils. Genetic analysis of the I. radicicola complex containing these pathogens was done by comparing the DNA sequences of the histone h3 region. These isolates originating from the rhizosphere soils of chestnut constituted a clade with other closely related species or I. radicicola isolates originating from ginseng or other host plants, respectively. Additionally, the pathogenicity tests to analyze the characteristics of these I. radicicola isolates revealed that they caused weakly virulent root rot on ginseng. Conclusions: This is the first study reporting that I. radicicola isolates from chestnut rhizosphere soils can attack ginseng plant in Korea. Thus, these results are expected to provide informations in the selection of suitable fields for ginseng cultivation.

Genetic Diversity of Taenia asiatica from Thailand and Other Geographical Locations as Revealed by Cytochrome c Oxidase Subunit 1 Sequences

  • Anantaphruti, Malinee Thairungroj;Thaenkham, Urusa;Watthanakulpanich, Dorn;Phuphisut, Orawan;Maipanich, Wanna;Yoonuan, Tippayarat;Nuamtanong, Supaporn;Pubampen, Somjit;Sanguankiat, Surapol
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.55-59
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    • 2013
  • Twelve 924 bp cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA sequences from Taenia asiatica isolates from Thailand were aligned and compared with multiple sequence isolates from Thailand and 6 other countries from the GenBank database. The genetic divergence of T. asiatica was also compared with Taenia saginata database sequences from 6 different countries in Asia, including Thailand, and 3 countries from other continents. The results showed that there were minor genetic variations within T. asiatica species, while high intraspecies variation was found in T. saginata. There were only 2 haplotypes and 1 polymorphic site found in T. asiatica, but 8 haplotypes and 9 polymorphic sites in T.saginata. Haplotype diversity was very low, 0.067, in T. asiatica and high, 0.700, in T. saginata. The very low genetic diversity suggested that T. asiatica may be at a risk due to the loss of potential adaptive alleles, resulting in reduced viability and decreased responses to environmental changes, which may endanger the species.

Generation of Transgenic Mice Overexpression Mouse RESISTIN

  • J. R. Chun;S. J. Song;J. T. Do;K. S. Chung;Lee, H. T.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.99-99
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    • 2002
  • The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

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Distribution and Bionomics of the Argentine Ant Linepithema humile (Mayr) (Hymenoptera: Formicidae: Dolichoderinae)

  • Min-Ji Lee;Young-Gyu Ban;Heejo Lee;Young Ha Kim;Dayeong Kim;Nang-Hee Kim;Dong Eon, Kim
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.4 no.3
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    • pp.104-114
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    • 2023
  • In this study, the distribution, bait preference, hosts, symbionts, habitat status, and cytochrome c oxidase subunit-I sequences of the ant Linepithema humile were analyzed. This species habitat comprised approximately 1.5 km along stone crevices, flower gardens, roadsides, and container yards in the vicinity of Busan station, Korea. Feeding activity of this species was confirmed in 9 species of 7 families of plants. Which was approximately 14% of total flora (63 species of 37 families) in the studied area. Particularly, it was observed that Dendranthema indicum and Camellia japonica were the most frequently visited. Further, we verified that they interacted with four species of aphids. We identified 22 species of ants (17 genera, 4 subfamilies) inhabiting the Busan station area, including L. humile, Tetramorium tsushimae, Nylanderia flavipes and the alien species Paratrechina longicornis. In areas where L. humile was the dominant species, the habitat of local native ants was reduced by approximately 30%. Bait preference was ranked in the following order: jelly, water with glucose (25%), and yogurt, with the highest preference for baits containing sugar and protein. The 630-bp cytochrome c oxidase subunit-I sequences of the local L. humile populations were 99% identical to those of L. humile in the National Center for Biotechnology Information database, and the Argentine ants that invaded Korea had a sequence identical to that of other invasive populations from China, Japan, and New Zealand. As L. humile rapidly increases due to domestic adaptation, continuous monitoring and control strategy for eradication are needed to protect domestic biodiversity.

Complete Genome Sequence of Massilia sp. KACC 81254BP Reveals a Potential for Degrading Polyhydroxyalkanoates

  • Sihyun An;Gyeongjun Cho;Jae-Hyung Ahn;Hang-Yeon Weon;Dayeon Kim;Young-Joon Ko;Jehyeong Yeon;Joon-hui Chung;Han Suk Choi;Jun Heo
    • Microbiology and Biotechnology Letters
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    • v.52 no.1
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    • pp.102-104
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    • 2024
  • Massilia sp. KACC 81254BP, isolated from a landfill on Jeju Island, Republic of Korea, possesses the capability to degrade polyhydroxyalkanoates (PHAs). The genomic analysis of strain KACC 81254BP consists of a circular chromosome comprising 5,028,452 base pairs with a DNA G+C content of 64.6%. This complete genome consists of a total of 4,513 genes, including those encoding the PHA degradation enzyme (PhaZ). This study offers valuable genomic insights into the enzymes responsible for degrading polyester plastics.

Mutations in the gyrB, parC, and parE Genes of Quinolone-Resistant Isolates and Mutants of Edwardsiella tarda

  • Kim, Myoung-Sug;Jun, Lyu-Jin;Shin, Soon-Bum;Park, Myoung-Ae;Jung, Sung-Hee;Kim, Kwang-Il;Moon, Kyung-Ho;Jeong, Hyun-Do
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1735-1743
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    • 2010
  • The full-length genes gyrB (2,415 bp), parC (2,277 bp), and parE (1,896 bp) in Edwardsiella tarda were cloned by PCR with degenerate primers based on the sequence of the respective quinolone resistance-determining region (QRDR), followed by elongation of 5' and 3' ends using cassette ligation-mediated PCR (CLMP). Analysis of the cloned genes revealed open reading frames (ORFs) encoding proteins of 804 (GyrB), 758 (ParC), and 631 (ParE) amino acids with conserved gyrase/topoisomerase features and motifs important for enzymatic function. The ORFs were preceded by putative promoters, ribosome binding sites, and inverted repeats with the potential to form cruciform structures for binding of DNA-binding proteins. When comparing the deduced amino acid sequences of E. tarda GyrB, ParC, and ParE with those of the corresponding proteins in other bacteria, they were found to be most closely related to Escherichia coli GyrB (87.6% identity), Klebsiella pneumoniae ParC (78.8% identity), and Salmonella Typhimurium ParE (89.5% identity), respectively. The two topoisomerase genes, parC and parE, were found to be contiguous on the E. tarda chromosome. All 18 quinolone-resistant isolates obtained from Korea thus far did not contain subunit alternations apart from a substitution in GyrA (Ser83$\rightarrow$Arg). However, an alteration in the QRDR of ParC (Ser84$\rightarrow$Ile) following an amino acid substitution in GyrA (Asp87$\rightarrow$Gly) was detected in E. tarda mutants selected in vitro at $8{\mu}g/ml$ ciprofloxacin (CIP). A mutant with a GyrB (Ser464$\rightarrow$Leu) and GyrA (Asp87$\rightarrow$Gly) substitution did not show a significant increase in the minimum inhibitory concentration (MIC) of CIP. None of the in vitro mutants exhibited mutations in parE. Thus, gyrA and parC should be considered to be the primary and secondary targets, respectively, of quinolones in E. tarda.

Cloning of Low-molecular-weight Glutenin Subunit Genes and Identification of their Protein Products in Common Wheat (Triticum aestivum L.) (보통 밀에서 저분자글루테닌 유전자 클로닝 및 단백질 동정)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kim, Bo-Mi;Lee, Jung-Hye;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Nam, Myung-Hee;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.547-554
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    • 2010
  • Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

Fine Mapping of the Rice Bph1 Gene, which Confers Resistance to the Brown Planthopper (Nilaparvata lugens Stal), and Development of STS Markers for Marker-assisted Selection

  • Cha, Young-Soon;Ji, Hyeonso;Yun, Doh-Won;Ahn, Byoung-Ohg;Lee, Myung Chul;Suh, Seok-Cheol;Lee, Chun Seok;Ahn, Eok Keun;Jeon, Yong-Hee;Jin, Il-Doo;Sohn, Jae-Keun;Koh, Hee-Jong;Eun, Moo-Young
    • Molecules and Cells
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    • v.26 no.2
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    • pp.146-151
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    • 2008
  • The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.