• Title/Summary/Keyword: N-terminal analysis

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Genetic Diversity of a Natural Population of Apple stem pitting virus Isolated from Apple in Korea

  • Yoon, Ju Yeon;Joa, Jae Ho;Choi, Kyung San;Do, Ki Seck;Lim, Han Cheol;Chung, Bong Nam
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.195-199
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    • 2014
  • Apple stem pitting virus (ASPV), of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP) gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD) and single breakpoint recombination (SBP) analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS) below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

Design and Performance Analysis of an Enhanced MAC Algorithm for the IEEE 802.11 DCF (IEEE 802.11 DCF 성능 개선을 위한 매체접근제어 알고리즘의 설계 및 성능 분석)

  • Hwang, An-Kyu;Lee, Jae-Yong;Kim, Byung-Chul
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.42 no.10 s.340
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    • pp.39-50
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    • 2005
  • In this paper, we propose a performance improving MAC algerian for the IEEE 802.11 DCF. WLAN based IEEE 802.11 uses two control methods called 'Distributed Coordination Function(UF)' and 'Point Coordination Function(PCF)'. The nF controls the Urnsmission based on carrier sense multiple access with collision detection(CSMA/CA), that decides a random backoff time with the range of contention window for each terminal. Normally, each terminal the CW double after collision, and reduces the CW to the minimum after successful transmission. This paper proposes an enhanced DCF algorithm that decreases the CW smoothly after successful transmission in order to reduce the collision Probability by utilizing the current status information of WLAN. We also analyze the throughput and delay performance for the unsaturated case mathematically. Simulation results show that our algorithm enhances the saturation throughput of WLAN. They also coincide well with the analytical results.

Molecular Cloning of ATPase $\alpha$-Subunit Gene from Mitochondria of Korean Ginseng (Panu ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A. Meyer) ATPase $\alpha$-subunit 유전자의 Cloning)

  • Park, Ui-Sun;Choi, Kwan-Sam;Kim, Kab-Sig;Kim, Nam-Won;Choi, Kwang-Tae
    • Journal of Ginseng Research
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    • v.19 no.1
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    • pp.56-61
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    • 1995
  • Molecular cloning and restriction mapping on ATPase $\alpha$-subunit gene (atpA) were carried out to obtain genomic information concerned with the gene structure and organization in Korean ginseng mitochondria. Two different clones containing the homologous sequence of atpA gene were selected from SalI and PstI libraries of mitochondrial DNA (mtDNA) of Korean ginseng. The sizes of mtDNA fragments inserted in SalI and PstI clones were 3.4 kb and 13 kb, respectively. Southern blot analysis with [$^{32}P$] labelled Oenothera atPA gene probe showed that atpA gene sequence was located in 2.0 kb XkaI fragment in PstI clone and in 1.7 kb XbaI fragment in SalI clone. A partial sequening ascertained that the SalI clone included about 1.2 kb fragment from SalI restriction site to C-terminal sequence of this gene but about 0.3 kb N-terminal sequence of open reading frame was abscent. The PstI fragment was enough large to cover the full sequence of atpA gene. The same restriction pattern of the overlapped region suggests that both clones include the same fragment of atiA locus. Data of Southern blot analysis and partial nucleotide sequencing suggested that mtDNA of Korean ginseng has a single copy of atpA gene. Key words ATPase a-subunit, mitochondrial DNA, Panax ginseng.

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Molecular identification and expression analysis of bactericidal permeability-increasing protein/ LPS-binding protein (BPI/LBP) from Black rockfish Sebastes schlegeli

  • Kwon, Mun-Gyeong;Kim, Ju-Won;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Park, Chan-Il
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.323-334
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    • 2010
  • Bactericidal/permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) are important components of the mammalian innate defence system against Gram-negative infections. The BPI/LBP cDNA was identified from the black rockfish ConA/PMA or LPS stimulated leukocyte cDNA library. The full-length BR-BPI/LBP cDNA was 2118 bp long and contained an open reading frame (ORF) of 1422 bp that encoded 473 amino-acid residues. The 5' UTR had a length of 57 bp, and the 3' UTR 639 bp. The molecular weight and theoretical isoelectric point (pI) values were calculated 51.4 kDa and 9.72, respectively. Compared with other known BPI or BPI/LBP peptide sequences, the most conserved regions of the black rockfish BPI/LBP peptide were found to be the BPI1 N-terminal, BPI2 C-terminal domains and a LPS binding domain. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the BPI/LBP sequence of black rockfish and that of other teleosts. The black rockfish BPI/LBP gene was predominantly expressed in the PBLs, head kidney, trunk kidney and spleen. The expression of the black rockfish BPI/LBP molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following LPS stimulation, with a peak at 12 h post-stimulation.

Development of a novel endolysin, PanLys.1, for the specific inhibition of Peptostreptococcus anaerobius

  • Joonbeom Moon;Hanbeen Kim;Dongseok Lee;Jakyeom Seo
    • Animal Bioscience
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    • v.36 no.8
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    • pp.1285-1292
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    • 2023
  • Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn2+ or Zn2+ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

Molecular cloning, identification, transcriptional analysis, and silencing of enolase on the life cycle of Haemaphysalis longicornis (Acari, Ixodidae) tick

  • Md. Samiul Haque;Md. Khalesur Rahman;Mohammad Saiful Islam;Myung-Jo You
    • Parasites, Hosts and Diseases
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    • v.62 no.2
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    • pp.226-237
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    • 2024
  • Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase's role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies.

Characterization of Heterogeneous Interaction Behaviour in Ternary Mixtures by Dielectric Analysis: The H-Bonded Binary Polar Mixture in Non-Polar Solvent

  • Sengwa, R.J.;Madhvi;Sankhla, Sonu;Sharma, Shobha
    • Bulletin of the Korean Chemical Society
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    • v.27 no.5
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    • pp.718-724
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    • 2006
  • The heterogeneous association behaviour of various concentration binary mixtures of mono alkyl ethers of ethylene glycol with ethyl alcohol were investigated by dielectric measurement in benzene solutions over the entire concentration range at 25 ${^{\circ}C}$. The values of static dielectric constant $\epsilon_0$ of the mixtures were measured at 1 MHz using a four terminal dielectric liquid test fixture and precision LCR meter. The high frequency limiting dielectric constant $\epsilon_\infty$ values were determined by measurement of refractive index $n_D$ ($\epsilon_\infty\;=\;n_D\;^2$). The measured values of $\epsilon_0$ and $\epsilon_\infty$ were used to evaluate the values of excess dielectric constant $\epsilon^E$, effective Kirkwood correlation factor $g^{eff}$ and corrective correlation factor $g_f$ of the binary polar mixtures to obtain qualitative and quantitative information about the H-bond complex formation. The non-linear behaviour of the observed $\epsilon_0$ values of the polar molecules and their mixtures in benzene solvent confirms the variation in the associated structures with change in polar mixture constituents concentration and also by dilution in non-polar solvents. Appearance of the maximum in $\epsilon^E$ values at different concentration of the polar mixtures suggest the formation of stable adduct complex, which depends on the molecular size of the mono alkyl ethers of ethylene glycol. Further, the observed $\epsilon^E$ < 0 also confirms the heterogeneous H-bond complex formation reduces the effective number of dipoles in these polar binary mixtures. In benzene solutions these polar molecules shows the maximum reduce in effective number of dipoles at 50 percent dilutions. But ethyl alcohol rich binary polar mixtures in benzene solvent show the maximum reduce in effective number of dipoles in benzene rich solutions.

Cytogenetic Analysis of Reciprocal Hybrids Reveals a Robertsonian Translocation between Mud Loach ($Misgurnus$ $mizolepis$) and Cyprinid Loach ($M.$ $anguillicaudatus$) (미꾸라지($Misgurnus$ $mizolepis$)와 미꾸리($M.$ $anguillicaudatus$) 및 유도된 종간 잡종의 세포유전학적 연구)

  • Lee, Seung-Ki;Kim, Dong-Soo
    • Korean Journal of Ichthyology
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    • v.24 no.1
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    • pp.1-10
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    • 2012
  • Reciprocal hybrids between the mud loach ($Misgurnus$ $mizolepis$) and cyprinid loach ($M.$ $anguillicaudatus$) were produced by artificial fertilization. The chromosome number of mud loach was 2n=48, consisting of 12M+4SM+32A chromosomes. The cyprinid loach has 2n=50, consisting of 10M+4SM+36A chromosomes. The chromosome numbers of the diploid reciprocal hybrids were 2n=49, consisting of 11M+4SM+34A chromosomes. All the karyotypes documented in this study had the same arm number of 64. There was no evidence of chromosomal polymorphisms or sex-related heteromorphism. The cytogenetic traits of the hybrid genotypes were intermediate between those of the parent species. In all genotypes, the chromosomal NORs localized to the terminal short arms of the same metacentric chromosome pair. These results suggest that Robertsonian translocation occurred between metacentric chromosome 1 of mud loach and acrocentric chromosome of cyprinid loach.

Analysis of the spike glycoprotein gene and nonstructural protein gene of transmissible gastroenteritis virus using PCR and RFLP analysis (PCR과 RFLP분석을 이용한 transmissible gastroenteritis virus의 spike glycoprotein gene과 nonstructural protein gene의 분석)

  • Kwon, Hyuk-moo
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.627-633
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    • 1996
  • To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

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Mutational Analysis Elucidates the Role of Conserved 237 Arginine in 23S rRNA Methylation, Which is in the Concave Cleft Region of ErmSF (ErmSF에서 두 도메인 사이에 존재하는 잘 보존된 237번 아르지닌 잔기의 위치 지정 치환 변이의 효소 활성 검색을 통한 역할 규명)

  • Jin, Hyung Jong
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.105-111
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    • 2013
  • The Erm family of adenine-$N^6$ methyltransferases (MTases) is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Recently, it has been proposed that well conserved amino acids in ErnC' located in concave cleft between N-terminal 'catalytic' domain and C-terminal 'RNA-binding' domain interacts with substrate RNA. We carried out the site-directed mutagenesis and studied the function of the ErmSF R237 mutant in vitro and in vivo. R237 amino acid residue is located in the concave cleft between two domains. Furthermore this residue is very highly conserved in almost all the Erm family. Purified mutant protein exhibited only 51% enzyme activity compared to wild-type. Escherichia coli with R237A mutant protein compared to the wild-type protein expressing E. coli did not show any difference in its MIC (minimal inhibitory concentration) suggesting that even with lowered enzyme activity, mutant protein was able to efficiently methylate 23S rRNA to confer the resistance on E. coli expressing this protein. But this observation strongly suggests that R237 of ErmSF probably interacts with substrate RNA affecting enzyme activity significantly.