• 제목/요약/키워드: elongation factor-2(EF-2)

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Molecular Systematics of the Genus Megoura (Hemiptera: Aphididae) Using Mitochondrial and Nuclear DNA Sequences

  • Kim, Hyojoong;Lee, Seunghwan
    • Molecules and Cells
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    • 제25권4호
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    • pp.510-522
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    • 2008
  • To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

러시아 철갑상어(Acipenser gueldenstaedtii) 발생 시료의 RT-qPCR 분석을 위한 내재 대조군 유전자의 선정 (Evaluation of Candidate Housekeeping Genes for the Normalization of RT-qPCR Analysis using Developing Embryos and Prolarvae in Russian Sturgeon Acipenser gueldenstaedtii)

  • 남윤권;이상윤;김은정
    • 한국수산과학회지
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    • 제51권1호
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    • pp.95-106
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    • 2018
  • To evaluate appropriate reference genes for the normalization of quantitative reverse transcription PCR (RT-qPCR) data with embryonic and larval samples from Russian sturgeon Acipenser gueldenstaedtii, the expression stability of eight candidate housekeeping genes, including beta-actin (ACTB), elongation factor-1A (EF1A), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone 2A (H2A), ribosomal protein L5 (RPL5), ribosomal protein L7 (RPL7), succinate dehydrogenase (SDHA), and ubiquitin-conjugating enzyme E2 (UBE2A), were tested using embryonic samples from 12 developmental stages and larval samples from 11 ontogenic stages. Based on the stability rankings from three statistic software packages, geNorm, NormFinder, and BestKeeper, the expression stability of the embryonic subset was ranked as UBE2A>H2A>SDHA>GAPDH>RPL5>EF1A>ACTB>RPL7. On the other hand, the ranking in the larval subset was determined as UBE2A>GAPDH>SDHA>RPL5>RPL7>H2A>EF1A>AC TB. When the two subsets were combined, the overall ranking was UBE2A>SDHA>H2A>RPL5>GAPDH>EF1A>ACTB>RPL7. Taken together, our data suggest that UBE2A and SDHA are recommended as suitable references for developmental and ontogenic samples of this sturgeon species, whereas traditional housekeepers such as ACTB and GAPDH may not be suitable candidates.

형태적 특징 및 다좌위 염기서열 분석에 의한 산철쭉 모무늬병균 Sphaerulina azaleae 동정 (Identification of Sphaerulina azaleae on Korean Azalea in Korea Based on Morphological Characteristics and Multilocus Sequence Typing)

  • 최인영;최영준;이귀재;주호종;조성완;신현동
    • 한국균학회지
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    • 제48권3호
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    • pp.329-335
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    • 2020
  • 2008년부터 2017년도에 제주, 홍천 등에서 산철쭉에서 모무늬 증상을 나타내는 잎을 채집하였다. 산철쭉모무늬 증상은 빈번하게 잎에만 발생하여 식물의 관상가치를 떨어트리고 조기낙엽을 유발하였다. 변색부는 잎의 윗면에 작고 담갈색 내지 흑자색 점무늬가 먼저 나타나며, 잎의 세맥으로 경계가 구분되어 모무늬 또는 부정형의 증상을 나타냈다. 산철쭉에서 분리한 균주를 동정하고자 형태적 특징과 actin (Act), translation elongation factor 1-alpha (EF), internal transcribed spacer (ITS), 28S nrDNA (LSU), RNA polymerase II second largest subunit (RPB2) 염기서열을 분석하였다. 형태적 특징을 재확인한 염기서열 분석결과 Sp. azaleae와 99~100%의 상동성을 나타냈으며, 계통수를 작성하였을 때도 Sp. azaleae 계통군에 속하였다. 따라서 산철쭉에 모무늬 증상에 관여하는 곰팡이는 Sp. azaleae로 동정되었다.

Two new species of Trichoderma isolated from commercially grown oyster mushroom, Pleurotus ostreatus (oral)

  • Park, Myung-Soo;Seo, Geon-Sik;Bae, Kyung-Sook;Yu, Seung-Hun
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.127.1-127
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    • 2003
  • We describe two new Trichoderma species associated with oyster mushroom in Korea. Trichoderma green mould has been one of the most serious diseases of oyster mushroom in Korea. Of these the predominant species are two unrecorded species. We designed as Trichoderma sp. Korean type 1 (Th K1) and Trichoderma sp. Korean type 2 (Th K2), respectively. Th K1 and Th K2 can be distinguished from previously reported Trichoderma species as well as each other in morphological characteristics including growth rate at 35$^{\circ}C$, colony morphology, conidia shape and branch pattern of phialides. Sequence of the ITS region of rDNA, the protein coding translation elongation factor gene(EF-1${\alpha}$), and RNA polymeraseII (RPB2) not only clearly separated Trichoderma sp. Korean types from their closely related T. harzianum biotype but also distinguished them from each other. Analyses of the EF-1${\alpha}$ and RPB2 sequences were found to be more useful for establishing systematic relationships among Trichoderma isolates than those of the ITS sequence. Based on the results of morphological and molecular characteristics. We propose the two Trichoderma sp. Korean types as the new species

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Protein tRNA Mimicry in Translation Termination

  • Nakamura, Yoshikazu
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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    • pp.83-89
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    • 2001
  • Recent advances in the structural and molecular biology uncovered that a set of translation factors resembles a tRNA shape and, in one case, even mimics a tRNA function for deciphering the genetic :ode. Nature must have evolved this 'art' of molecular mimicry between protein and ribonucleic acid using different protein architectures to fulfill the requirement of a ribosome 'machine'. Termination of protein synthesis takes place on the ribosomes as a response to a stop, rather than a sense, codon in the 'decoding' site (A site). Translation termination requires two classes of polypeptide release factors (RFs): a class-I factor, codon-specific RFs (RFI and RF2 in prokaryotes; eRFI in eukaryotes), and a class-IT factor, non-specific RFs (RF3 in prokaryotes; eRF3 in eukaryotes) that bind guanine nucleotides and stimulate class-I RF activity. The underlying mechanism for translation termination represents a long-standing coding problem of considerable interest since it entails protein-RNA recognition instead of the well-understood codon-anticodon pairing during the mRNA-tRNA interaction. Molecular mimicry between protein and nucleic acid is a novel concept in biology, proposed in 1995 from three crystallographic discoveries, one, on protein-RNA mimicry, and the other two, on protein-DNA mimicry. Nyborg, Clark and colleagues have first described this concept when they solved the crystal structure of elongation factor EF- Tu:GTP:aminoacyl-tRNA ternary complex and found its overall structural similarity with another elongation factor EF-G including the resemblance of part of EF-G to the anticodon stem of tRNA (Nissen et al. 1995). Protein mimicry of DNA has been shown in the crystal structure of the uracil-DNA glycosylase-uracil glycosylase inhibitor protein complex (Mol et al. 1995; Savva and Pear 1995) as well as in the NMR structure of transcription factor TBP-TA $F_{II}$ 230 complex (Liu et al. 1998). Consistent with this discovery, functional mimicry of a major autoantigenic epitope of the human insulin receptor by RNA has been suggested (Doudna et al. 1995) but its nature of mimic is. still largely unknown. The milestone of functional mimicry between protein and nucleic acid has been achieved by the discovery of 'peptide anticodon' that deciphers stop codons in mRNA (Ito et al. 2000). It is surprising that it took 4 decades since the discovery of the genetic code to figure out the basic mechanisms behind the deciphering of its 64 codons.

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Phosphoinositide 3-kinase regulates myogenin expression at both the transcriptional and post-transcriptional level during myogenesis

  • Woo, Joo-Hong;Kim, Min-Jeong;Kim, Hye-Sun
    • Animal cells and systems
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    • 제14권3호
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    • pp.147-154
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    • 2010
  • It is well-established that phosphoinositide 3-kinase (PI3-kinase) regulates myogenesis by inducing transcription of myogenin, a key muscle regulatory factor, at the initiation of myoblast differentiation. In this study, we investigated the role of PI3-kinase in cells that have committed to differentiation. PI3-kinase activity increases during myogenesis, and this increase is sustained during the myogenic process; however, its function after the induction of differentiation has not been investigated. We show that LY294002, a PI3-kinase inhibitor, blocked myoblast fusion even after myogenin expression initially increased. In contrast to the inhibitory effects of LY294002 on myogenin mRNA levels during the initiation of differentiation, LY294002 blocked the accumulation of myogenin protein without affecting its mRNA level after differentiation was induced. Treatment with cycloheximide, a translation inhibitor, or actinomycin D, a transcription inhibitor, indicated that the stability of myogenin protein is lower than that of its mRNA. LY294002 inhibited the activities of several important translation factors, including eukaryotic elongation factor-2(eEF2), by altering their phosphorylation status. In addition, LY294002 blocked the incorporation of [$^{35}S$]methionine into newly synthesized proteins. Since myogenin has a relatively short half-life, LY294002-mediated inhibition of post-transcriptional processes resulted in a rapid depletion of myogenin protein. In summary, these results suggest that PI3-kinase plays an important role in regulating the expression of myogenin through post-transcriptional mechanisms after differentiation has been induced.

수확기 벼 이삭에서 분리된 진균독소 생성 Fusarium armeniacum의 다양성 (Diversity of Mycotoxigenic Fusarium armeniacum Isolated from Rice Grains at Harvest Time in Korea)

  • 홍성기;이수형;이데레사;함현희;문혜연;최효원;손승완;류재기
    • 한국균학회지
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    • 제43권3호
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    • pp.158-164
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    • 2015
  • 2010년부터 2015년까지 전국 8도에 있는 벼 포장에서 수확기에 총 509개의 이삭시료가 채집되었다. 시료당 105개의 벼 종실이 potato dextrose agar (PDA) 배지에 처리되었고 6,658개의 Fusarium 균주가 분리되었다. $EF-1{\alpha}$ 유전자의 염기서열을 기초로, 분리된 Fusarium 중 67균주를 Fusarium armeniacum으로 동정하였고, 형태적, 배양적 특성을 확인하였다. F. armeniacum은 분생포자 크기, 균총 색, 및 $EF-1{\alpha}$ 염기서열에서 균주간에 상당한 차이가 있었다. 액체크로마토크래피-질량분석기를 사용하여 potato sucrose agar (PSA) 배지에서 T-2와 HT-2 독소생성능력을 결정하였던 바 F. armeniacum 24균주 중 21균주가 T-2와 HT-2 독소를 모두 생성하였으며, 독소생성 수준은 균주간에 다양하였다. 이러한 결과는 한국산 F. armeniacum 균주들이 형태적, 배양적, 유전적 및 독소학적 성질에서 다양성을 갖는다는 것을 보여준다.

Molecular Diagnosis of Taenia saginata Tapeworms from Two Residents of Northern Cambodia

  • Chang, Taehee;Jung, Bong-Kwang;Sohn, Woon-Mok;Hong, Sooji;Shin, Hyejoo;Ryoo, Seungwan;Lee, Jeonggyu;Lee, Keon Hoon;Khieu, Virak;Huy, Rekol;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제58권2호
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    • pp.201-204
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    • 2020
  • Taenia saginata infection has seldom been reported in Cambodia. In this study, we performed a survey of intestinal parasites in 1,156 residents of Preah Vihear and Stung Treng Provinces in 2018. The results revealed that 26 (2.4%) cases were positive for Taenia spp. eggs. In order to obtain the strobilae of the tapeworms, 2 patients in Preah Vihear were treated with praziquantel and purged with magnesium salts. The proglottids expelled after the medication were morphologically and molecularly analyzed to determine the species. The main uterine lateral braches in gravid proglottids were > 15 in number suggesting that they are either T. saginata or Taenia asiatica. The sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and 2 nuclear loci, elongation factor-1 alpha (ef1) and ezrin-radixin-moesin-like protein (elp), were identical to the sequences of T. saginata available in GenBank but distant from Taenia solium, T. asiatica, and T. saginata-T. asiatica hybrid. This is the first report of the presence of T. saginata in the northern part of Cambodia bordering Lao PDR based on a molecular confirmation.

EGCG Blocked Phenylephrin-Induced Hypertrophy in H9C2 Cardiomyocytes, by Activating AMPK-Dependent Pathway

  • Cai, Yi;Zhao, Li;Qin, Yuan;Wu, Xiao-Qian
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권3호
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    • pp.203-210
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    • 2015
  • AMP-activated protein kinase (AMPK) is a key regulator of energy metabolism. Previous studies have shown that activation of AMPK results in suppression of cardiac myocyte hypertrophy via inhibition of the p70S6 kinase (p70S6K) and eukaryotic elongation factor-2 (eEF2) signaling pathways. Epigallocatechin-3-gallate (EGCG), the major polyphenol found in green tea, possesses multiple protective effects on the cardiovascular system including cardiac hypertrophy. However, the molecular mechanisms has not been well investigated. In this study, we found that EGCG could significantly reduce natriuretic peptides type A (Nppa), brain natriuretic polypeptide (BNP) mRNA expression and decrease cell surface area in H9C2 cardiomyocytes stimulated with phenylephrine (PE). Moreover, we showed that AMPK is activated in H9C2 cardiomyocytes by EGCG, and AMPK-dependent pathway participates in the inhibitory effects of EGCG on cardiac hypertrophy. Taken together, our findings provide the first evidence that the effect of EGCG against cardiac hypertrophy may be attributed to its activation on AMPK-dependent signaling pathway, suggesting the therapeutic potential of EGCG on the prevention of cardiac remodeling in patients with pressure overload hypertrophy.

Occurrence of a Hybrid Between Taenia saginata and Taenia asiatica Tapeworms in Cambodia

  • Chang, Taehee;Jung, Bong-Kwang;Hong, Sooji;Shin, Hyejoo;Ryoo, Seungwan;Lee, Jeonggyu;Lee, Keon Hoon;Park, Hansol;Eom, Keeseon S.;Khieu, Virak;Huy, Rekol;Sohn, Woon-Mok;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제59권2호
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    • pp.179-182
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    • 2021
  • Human infection with Taenia asiatica or a hybrid between Taenia saginata and T. asiatica has not been reported in Cambodia. We detected for the first time a hybrid form between T. saginata and T. asiatica in Preah Vihear Province, Cambodia. An adult tapeworm specimen, i.e., 75 cm long strobila without scolex, was expelled from a 27-year-old man after praziquantel medication and purging. It was morphologically indistinguishable between T. saginata and T. asiatica. Several proglottids were molecularly analyzed to confirm the tapeworm species. The mitochondrial gene encoding cytochrome c oxidase subunit 1 (cox1) and nuclear genes encoding elongation factor-1α (ef1) and ezrin-radixin-moesin (ERM)-like protein (elp) were sequenced, and a single-allele analysis was performed to confirm the haploid genotype. The results revealed that our sample showed a discrepancy between the mitochondrial and 2 nuclear genes. It possessed homozygous sequences typical of T. saginata at cox1 and ef1 loci. However, it was heterozygous at the elp locus, with 1 allele in T. asiatica (elpA) and 1 in T. saginata (elpC), which indicates that it is a hybrid between T. saginata and T. asiatica. The present results confirmed the presence of a hybrid between T. saginata and T. asiatica in Cambodia and strongly suggest the existence of also 'pure' T. asiatica in Cambodia.