• Title/Summary/Keyword: that-Deletion

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A Brucella Omp16 Conditional Deletion Strain Is Attenuated in BALB/c Mice

  • Zhi, Feijie;Fang, Jiaoyang;Zheng, Weifang;Li, Junmei;Zhang, Guangdong;Zhou, Dong;Jin, Yaping;Wang, Aihua
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.6-14
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    • 2022
  • Brucella spp. are facultative intracellular pathogens that invade, survive and proliferate in numerous phagocytic and non-phagocytic cell types, thereby leading to human and animal brucellosis. Outer membrane proteins (Omps) are major immunogenic and protective antigens that are implicated in Brucella virulence. A strain deleted of the omp16 gene has not been obtained which suggests that the Omp16 protein is vital for Brucella survival. Nevertheless, we previously constructed an omp16 conditional deletion strain of Brucella, ∆Omp16. Here, the virulence and immune response elicted by this strain were assessed in a mouse model of infection. Splenomegaly was significantly reduced at two weeks post-infection in ∆Omp16-infected mice compared to infection with the parental strain. The bacterial load in the spleen also was significantly decreased at this post-infection time point in ∆Omp16-infected mice. Histopathological changes in the spleen were observed via hematoxylin-eosin staining and microscopic examination which showed that infection with the ∆Omp16 strain alleviated spleen histopathological alterations compared to mice infected with the parental strain. Moreover, the levels of humoral and cellular immunity were similar in both ∆Omp16-infected mice and parental strain-infected mice. The results overall show that the virulence of ∆Omp16 is attenuated markedly, but that the immune responses mediated by the deletion and parental strains in mice are indistinguishable. The data provide important insights that illuminate the pathogenic strategies adopted by Brucella.

Multiple Sluicing in English

  • Park, Myung-Kwan;Kang, Jung-Min
    • Proceedings of the Korean Society for Language and Information Conference
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    • 2007.11a
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    • pp.394-404
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    • 2007
  • This paper explores the nature of multiple sluicing in English, which has two or more remnant wh-phrases in clause edge position. At the beginning part of the paper we argue against Nishigauchi's (1998) and Lasnik's (2007) Gapping analysis of multiple sluicing, which says that two remnant wh-phrases each actually occupies the left and right edge of a clause, with the in-between string of words undergoing Gapping. We rather argue that multiple sluicing in English is the same kind as found in Bulgarian and Serbo-Croatian. In other words, multiple sluicing in English is also derived by multiple wh-fronting which otherwise does not apply. We demonstrate that some important properties of the construction noted by Lmultiple sluicing, multiple wh-movement/fronting, sluicing, TP/IP-deletion, asnik (2007) under the Gapping approach to it can be accounted for in a principled way by our proposed analysis.

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Isolation and characterization of the rice NPR1 promoter

  • Hwang, Seon-Hee;Hwang, Duk-Ju
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.29-35
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    • 2010
  • NPR1 is a positive regulator of systemic acquired resistance in Arabidopsis and rice. Expression of the rice gene OsNPR1 is induced by salicylic acid (SA). To identify the region of the OsNPR1 promoter involved in response to SA, we carried out deletion mutagenesis of the region 1005 bp upstream of the OsNPR1 start codon. Ciselement analysis revealed that the OsNPR1 promoter contains W-boxes and ASF1 motifs, both of which are known to be functional cis-elements of the WRKY and bZIP proteins, respectively. The deletion constructs 1005:LUC and 752:LUC, were induced by up to 4.3- and 3.8-fold, respectively, following SA treatment, suggesting that W-boxes and ASF1 motifs may play an important role in the strong induction of these constructs by SA. Using mutation analysis, we also showed that both the W-box and ASF1 motif are necessary for SA-induced expression of OsNPR1.

Role of 5'-UTR hairpins of the Turnip yellow mosaic virus RNA in replication and systemic movement

  • Shin, Hyun-Il;Cho, Nam-Jeong;Cho, Tae-Ju
    • BMB Reports
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    • v.41 no.11
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    • pp.778-783
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    • 2008
  • Turnip yellow mosaic virus (TYMV) RNA has two hairpins in its 5' untranslated region (5'-UTR). To investigate the role of the hairpins in replication of TYMV, mutants lacking one or both of the two hairpins were constructed. The TYMV constructs were introduced into Chinese cabbage by an Agrobacterium-mediated T-DNA transfer method, called agroinfiltration. Analysis of total RNA from agroinfiltrated leaves showed that replication of the mutant TYMV RNA lacking both hairpins was about 1/100 of wild type. This mutant was also impaired in systemic spread. Deletion analysis of each hairpin revealed that both hairpins were needed for maximal replication. The deletion analysis along with sequence modification of the hairpin structure indicates that the second hairpin plays a role in efficient long-distance systemic movement of TYMV.

Construction and Characterization of a Burkholderia pseudomallei wzm Deletion Mutant

  • Yuen, Chee-Wah;Ong, Eugene Boon Beng;Mohamad, Suriani;Manaf, Uyub Abdul;Najimudin, Nazalan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1336-1342
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    • 2012
  • In Burkholderia pseudomallei, the pathogen that causes melioidosis, the gene cluster encoding the capsular polysaccharide, is located on chromosome 1. Among the 19 capsular genes in this cluster, wzm has not been thoroughly studied. To study the function of wzm, we generated a deletion mutant and compared it with the wild-type strain. The mutant produced less biofilm in minimal media and was more sensitive to desiccation and oxidative stress compared with the wild-type strain, indicating that wzm is involved in biofilm formation and membrane integrity. Scanning electron microscopy showed that the bacterial cells of the mutant strain have more defined surfaces with indentations, whereas cells of the wild-type strain do not.

Analysis of Human O-GlcNAcase Gene and the Expression of the Recombinant Gene. (사람의 O-linked N-acetyl-$\beta$-D-glucosaminidase 유전자의 분석과 재조합 발현)

  • 강대욱;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.87-93
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    • 2004
  • Dynamic modification of cytoplasmic and nuclear proteins by O-linked N-acetylglucosamine (O-GlcNAc) on Ser and Thr residues is ubiquitous in higher eukaryotes. And this modification may serve as a signaling mod-ification analogous to protein phosphorylation. Addition and cleavage of O-GlcNAc are catalyzed by O-linked GlcNAc transferase (OGT) and O-linked N-acety1glucosaminidase (O-GlcNAcase), respectively. Two types of human O-GlcNAcase gene were cloned and expressed as three fusion proteins in Escherichia coli. O-GlcNA-case activity showed in the order of thioredoxin fusion> $6{\times}His$ tag> GST fusion. O-GlcNAcase had enzy-matic activity against only ${\rho}$NP-GlcNAc of seven tested substrate analogs. Blast search revealed that O-GlcNAcase has two conserved domains, amino terminal hyaluronidase-like domain and carboxy terminal N-acetyltransferase domain. Extensive deletion studies were done to define catalytically important domains. The deletions of hyaluronidase-like domain and N-acetyltransferase domain abolished enzyme activity. But, N-ter-minal 55 amino acid deletion and C-terminal truncation showed lower activity. Based on deletion analysis, we suggest that hyaluronidase-like domain is essential for enzyme activity and carboxy terminal N-acetyltrans-ferase domain may be modulatory function.

Domain Function and Relevant Enzyme Activity of Cycloinulooligosaccharide Fructanotransferase from Paenibacillus polymyxa (Paenibacillus polymyxa Cycloinulooligosaccharide Fructanotransferase의 효소 활성에 미치는 각 Domain의 역할)

  • You Dong-Ju;Park Jung-Ha;You Kyung-Ok;Nam Soo-Wan;Kim Kwang-Hyeon;Kim Byung-Woo;Kwon Hyun-Ju
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.278-287
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    • 2006
  • Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cycloinulooligosaccharides (cyclofructan, CF) of ${\beta}-(2{\to}1)$-linked D-fructofuranose as well as hydrolysis of cyclofructan. Sequences analysis indicated that CFTase was divided into five distinct regions containing three repeated sequences (R1, R3, and R4) at the N-terminus and C-terminus. Each domain function was investigated by comparison of wild type CFTase enzyme (CFT148) and deletion mutant proteins (CFT108: R1 and R3 deletion; CFT130: R4 deletion; and CFT88: R1, R3, and R4 deletion) of CFTase. The CFT108 mutant had both CFTase and CF hydrolyzing activity as CFT148 did. CFTase activities and CF hydrolysing activities were disappeared in CFT130 and CFT88 mutants. These results indicated that the C-terminal R4 region of P. polymyxa CFTase is necessary for cyclization and hydrolyzing activity.

Research on the Bottom Boundary Line on the Southeast Area of the Chungcheongdo Dialect in Yeongdong (영동지역어내의 충청방언 남동부 하한선 연구)

  • Seong, Hee-Jae
    • Lingua Humanitatis
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    • v.8
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    • pp.265-289
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    • 2006
  • The geographical characteristics of Yeongdong(永同) the southernmost part of the Chungcheongbukdo province, has attracted attention among the academic circle as one of the dialectal contact regions since it adjoins the Gyeongsang and Jeolla dialects. Unlike the local language in Mooju (Jellado dialect) adjacent to the Southwest part, the local language in Yeongdong is quite different from that of Kimcheon (Gyeongsang dialect). More specifically, it is noteworthy that the boundary line of the Gyeongsang dialect is found in this region, which is different from the administrative division. In other words, the local language in Yeongdong is divided into the Chungcheong dialect and the Gyeongsang dialect, and furthermore each dialect region still has the characteristics of the other region's dialect. For example, the phonological structure of Yeongdong Chungcheongdo dialect has very unique characteristics of the fudged dialect, which is seemingly influenced by the Gyeongsang dialect. The present study is to define the bottom boundary line of the southeast area of the Chungcheong dialect by identifying the boundary line between the Gyeongsang dialect and the Chungcheong dialect, and to clarify its specific sound system generated by the contact of these two dialects. For this, the author collected and analyzed data of the local language around Yeongdong and adjacent areas. It was found that Cheongwha-ri, Deokjin-ri, and Sanjeo-ri at Yeongsan-myeon, and Mugeunjeom, Sangga-ri, and Jungga-ri at Yeongdong-eup, among the regions that belongs to Chungcheong dialect within the local language of Yeongdong, show the characteristics of the Gyeongsang dialect. Accordingly, the western areas of these villages become the southeast boundary line of the Chungcheong dialect. Also, the unique phonological characteristics of the Yeongdong Chungcheong dialect is affected by the Gyeongsang dialect, among which "rhythms, y deletion, nasal phoneme deletion, and w deletion" appeared. It is thought to be the unique fudged dialectal phenomenon that appeared only in this region. The research result is expected to be of some help in finding out various aspects of dialectal contacts as well as clarifying the phonological features of the local language in Yeongdong, and thereby contributing to exact divisioning of the Chungcheong dialect.

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Regulation of amyloid precursor protein processing by its KFERQ motif

  • Park, Ji-Seon;Kim, Dong-Hou;Yoon, Seung-Yong
    • BMB Reports
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    • v.49 no.6
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    • pp.337-343
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    • 2016
  • Understanding of trafficking, processing, and degradation mechanisms of amyloid precursor protein (APP) is important because APP can be processed to produce β-amyloid (Aβ), a key pathogenic molecule in Alzheimer's disease (AD). Here, we found that APP contains KFERQ motif at its C-terminus, a consensus sequence for chaperone-mediated autophagy (CMA) or microautophagy which are another types of autophagy for degradation of pathogenic molecules in neurodegenerative diseases. Deletion of KFERQ in APP increased C-terminal fragments (CTFs) and secreted N-terminal fragments of APP and kept it away from lysosomes. KFERQ deletion did not abolish the interaction of APP or its cleaved products with heat shock cognate protein 70 (Hsc70), a protein necessary for CMA or microautophagy. These findings suggest that KFERQ motif is important for normal processing and degradation of APP to preclude the accumulation of APP-CTFs although it may not be important for CMA or microautophagy.

Conserved Roles of MonA in Fungal Growth and Development in Aspergillus Species

  • Son, Ye-Eun;Park, Hee-Soo
    • Mycobiology
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    • v.47 no.4
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    • pp.457-465
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    • 2019
  • MonA is a subunit of a guanine nucleotide exchange factor that is important for vacuole passing and autophagy processes in eukaryotes. In this study, we characterized the function of MonA, an orthologue of Saccharomyces cerevisiae Mon1, in the model fungus Aspergillus nidulans and a toxigenic fungus A. flavus. In A. nidulans, the absence of AnimonA led to decreased fungal growth, reduced asexual reproduction, and defective cleistothecia production. In addition, AnimonA deletion mutants exhibited decreased spore viability, had reduced trehalose contents in conidia, and were sensitive to thermal stress. In A. flavus, deletion of AflmonA caused decreased fungal growth and defective production of asexual spores and sclerotia structures. Moreover, the absence of monA affected vacuole morphology in both species. Taken together, these results indicate that MonA plays conserved roles in controlling fungal growth, development and vacuole morphology in A. nidulans and A. flavus.