• Title/Summary/Keyword: Deletion

Search Result 1,555, Processing Time 0.027 seconds

A Novel Oxidative Stress-inducible Peroxidase Promoter and Its Applications to Production of Pharmaceutical Proteins in Transgenic Cell Cultures

  • Lee, Ok-Sun;Park, Sun-Mi;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kim, Kee-Yeun;Kim, Jae-Whune;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
    • /
    • v.4 no.4
    • /
    • pp.143-150
    • /
    • 2002
  • A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

A Logit Type of Public Transit Trip Assignment Model Considering Stepwise Transfer Coefficients (Stepwise 환승계수를 고려한 Logit 유형 대중교통통행배정모형)

  • SHIN, Seongil;BAIK, Namcheol
    • Journal of Korean Society of Transportation
    • /
    • v.34 no.6
    • /
    • pp.570-579
    • /
    • 2016
  • This study proposes a concept of Stepwise Transfer Coefficient(STC) which implies greater transfer cost with increasing the number of transfers. Thus, the public transport information system provides the choice sets of travel routes by the consideration of not only transportation time but also the optimum number of transfers. However, path choice problems that involve STC are found to include non additive cost, which requires additional route enumeration works. Discussions on route enumeration in actual transportation networks is very complicated, thereby warranting a theoretical examination of route search considering STC. From these points of view, this study results in a probability based transit trip assignment model including STC. This research also uses incoming link based entire route deletion method. The entire route deletion method proposed herein simplifies construction of an aggregation of possible routes by theoretically supporting the process of enumeration of the different routes from origin to destination. Conclusively, the STC reflected route based logit model is proposed as a public transportation transit trip assignment model.

Functional Roles of a Putative B' Delta Regulatory Subunit and a Catalytic Subunit of Protein Phosphatase 2A in the Cereal Pathogen Fusarium graminearum

  • Kim, Hee-Kyoung;Yun, Sung-Hwan
    • The Plant Pathology Journal
    • /
    • v.28 no.3
    • /
    • pp.259-269
    • /
    • 2012
  • Protein phosphatase 2A (PP2A), a family of serine/threonine protein phosphatases, plays an important role in balancing the phosphorylation status of cellular proteins for regulating diverse biological functions in eukaryotic organisms. Despite intensive studies in mammals, limited information on its role is available in filamentous fungi. Here, we investigated the functional roles of genes for a putative B' delta regulatory subunit (FgPP2AR) and a catalytic subunit (FgPP2AC) of PP2A in a filamentous ascomycete, Fusarium graminearum. Molecular characterization of an insertional mutant of this plant pathogenic fungus allowed us to identify the roles of FgPP2AR. Targeted gene replacement and complementation analyses demonstrated that the deletion of FgPP2AR, which was constitutively expressed in all growth stages, caused drastic changes in hyphal growth, conidia morphology/germination, gene expression for mycotoxin production, sexual development and pathogenicity. In particular, overproduction of aberrant cylindrical-shaped conidia is suggestive of arthroconidial induction in the ${\Delta}FgPP2AR$ strain, which has never been described in F. graminearum. In contrast, the ${\Delta}FgPP2AC$ strain was not significantly different from its wild-type progenitor in conidiation, trichothecene gene expression, and pathogenicity; however, it showed reduced hyphal growth and no perithecial formation. The double-deletion ${\Delta}FgPP2AR;{\Delta}FgPP2AC$ strain had more severe defects than single-deletion strains in all examined phenotypes. Taken together, our results indicate that both the putative regulatory and catalytic subunits of PP2A are involved in various cellular processes for fungal development in F. graminearum.

Molecular Identification of Lipase LipA from Pseudomonas protegens Pf-5 and Characterization of Two Whole-Cell Biocatalysts Pf-5 and Top10lipA

  • Zha, Daiming;Xu, Li;Zhang, Houjin;Yan, Yunjun
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.5
    • /
    • pp.619-628
    • /
    • 2014
  • To identify lipase LipA (PFL_0617) from Pseudomonas protegens Pf-5, a lipA deletion mutant (Pf0617) and a complementary strain (Pf0617lipA) were constructed, and their effects on the lipase production were examined. Pf0617 remarkably decreased its whole-cell lipase activity, whereas Pf0617lipA made its whole-cell lipase activity not only restore to wild-type level but also get a further increment. However, the deletion and overexpression of lipA did not affect the extracellular lipase activity. In addition, the unbroken whole cells of these strains were able to catalyze the hydrolysis of membrane-permeable p-nitrophenyl esters, but could not hydrolyze the membrane-impermeable olive oil. These results confirmed that LipA was an intracellular lipase and Pf-5 could also be used as a natural whole-cell biocatalyst. To evaluate the potential of Pf-5 as a whole-cell biocatalyst and separately characterize the whole-cell LipA, the properties of the whole-cell lipases from Pf-5 and Top10lipA were characterized. The results demonstrated that both Pf-5 and Top10lipA exhibited high tolerance to alkaline condition, high temperature, heavy metal ions, surfactants, and organic solvents. Taken together, lipA can realize functional expression in E. coli Top10, and Pf-5 and Top10lipA as whole-cell biocatalysts may have enormous potential in applications.

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
    • /
    • v.8
    • /
    • pp.265-289
    • /
    • 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.

  • PDF

The Role of Abp140p in Actin Dynamics of Budding Yeast

  • Lim, Bum-Soon;Lee, Yong-Keun;Pon, Liza A.;Yang, Hyeong-Cheol
    • International Journal of Oral Biology
    • /
    • v.30 no.1
    • /
    • pp.17-22
    • /
    • 2005
  • In the previous studies of Saccharomyces cerevisiae, Abp140p (actin binding protein 140) fused to GFP has been only a protein that can label actin cables of yeast cells so far. However, the role of Abp140p in actin dynamics was remained elusive. In this study, the function of Abp140p was investigated with a deletion mutant and overexpression of GFP fused Abp140p. The deletion mutant was slightly more susceptible to Latrunculin-A (Lat-A), an actin-monomer sequestering agent, than wild type, although no significant deformation of actin structures was caused by ABP 140 deletion. Overexpression of Abp140p-GFP retarded cell growth, and produced thick and robust actin cables. Lat-A was not able to destabilize the thick actin cables, which suggests that actin dynamics was compromised in the cells with surplus of Abp140p. Therefore, Abp140p seems to stabilize actin cables together with other bundling proteins. Recently, actin cable dynamics of budding yeast was found to have a resemblance to that of filopodial tip of cultured mammalian cells. Retrograde movement of actin cables from buds to mother cells indicated local generation of the cable at bud sites. By using Abp140p-GFP, we traced the steps in the generation of a new actin cable after elimination of old cables by sodium azide. Before the appearance of a new actin cable, Abp140p-GFP concentrated in buds and disappeared, as mother cells became abundant in actin cables. Our observations provide a direct evidence of actin cable formation at buds of budding cells.

Inhibition of GM3 Synthase Attenuates Neuropathology of Niemann-Pick Disease Type C by Affecting Sphingolipid Metabolism

  • Lee, Hyun;Lee, Jong Kil;Bae, Yong Chul;Yang, Song Hyun;Okino, Nozomu;Schuchman, Edward H.;Yamashita, Tadashi;Bae, Jae-Sung;Jin, Hee Kyung
    • Molecules and Cells
    • /
    • v.37 no.2
    • /
    • pp.161-171
    • /
    • 2014
  • In several lysosomal storage disorders, including Niemann-Pick disease Type C (NP-C), sphingolipids, including glycosphingolipids, particularly gangliosides, are the predominant storage materials in the brain, raising the possibility that accumulation of these lipids may be involved in the NP-C neurodegenerative process. However, correlation of these accumulations and NP-C neuropathology has not been fully characterized. Here we derived NP-C mice with complete and partial deletion of the Siat9 (encoding GM3 synthase) gene in order to investigate the role of ganglioside in NP-C pathogenesis. According to our results, NP-C mice with homozygotic deletion of GM3 synthase exhibited an enhanced neuropathological phenotype and died significantly earlier than NP-C mice. Notably, in contrast to complete depletion, NP-C mice with partial deletion of the GM3 synthase gene showed ameliorated NP-C neuropathology, including motor disability, demyelination, and abnormal accumulation of cholesterol and sphingolipids. These findings indicate the crucial role of GM3 synthesis in the NP-C phenotype and progression of CNS pathologic abnormality, suggesting that well-controlled inhibition of GM3 synthesis could be used as a therapeutic strategy.

Fluorescence-in-situ-hybridization in the Surveillance of Urothelial Cancers: Can Use of Cystoscopy or Ureteroscopy be Deferred?

  • Ho, Christopher Chee Kong;Tan, Wei Phin;Pathmanathan, Rajadurai;Tan, Wei Keith;Tan, Hui Meng
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.7
    • /
    • pp.4057-4059
    • /
    • 2013
  • Background: Fluorescence in situ hybridization (FISH) testing may be useful to screen for bladder carcinoma or dysplasia by detecting aneuploidy chromosomes 3, 7, 17 and deletion of the chromosome 9p21 locus in urine specimens. This study aimed to assess the sensitivity, specificity, positive and negative predictive value of FISH in a multi-ethnic population in Asia. Materials and Methods: Patients with haematuria and/or past history of urothelial cancer on follow-up had their voided urine tested with FISH. Patients then underwent cystoscopy/ureteroscopy and any lesions seen were biopsied. The histopathological reports of the bladder or ureteroscopic mucosal biopsies were then compared with the FISH test results. Results: Two hundred sixty patients were recruited. The sensitivity and specificity of the FISH test was 89.2% and 83.4% respectively. The positive (PPV) and negative predictive values (NPV) were 47.1% and 97.9%. By excluding patients who had positive deletion of chromosome 9, the overall results of the screening test improved: sensitivity 84.6%; specificity 96.4%; PPV 75.9% and NPV 97.9%. Conclusions: UroVysion FISH has a high specificity of detecting urothelial cancer or dysplasia when deletion of chromosome 9 is excluded. Negative UroVysion FISH-tests may allow us to conserve health resources and minimize trauma by deferring cystoscopic or ureteroscopic examination.

Outlier Detection Based on MapReduce for Analyzing Big Data (대용량 데이터 분석을 위한 맵리듀스 기반의 이상치 탐지)

  • Hong, Yejin;Na, Eunhee;Jung, Yonghwan;Kim, Yangwoo
    • Journal of Internet Computing and Services
    • /
    • v.18 no.1
    • /
    • pp.27-35
    • /
    • 2017
  • In near future, IoT data is expected to be a major portion of Big Data. Moreover, sensor data is expected to be major portion of IoT data, and its' research is actively carried out currently. However, processed results may not be trusted and used if outlier data is included in the processing of sensor data. Therefore, method for detection and deletion of those outlier data before processing is studied in this paper. Moreover, we used Spark which is memory based distributed processing environment for fast processing of big sensor data. The detection and deletion of outlier data consist of four stages, and each stage is implemented with Mapper and Reducer operation. The proposed method is compared in three different processing environments, and it is expected that the outlier detection and deletion performance is best in the distributed Spark environment as data volume is increasing.

Clonal Analysis of Methicillin-Resistant Staphylococcus aureus Strains in Korea

  • Kim, Jung-Min;Seol, Sung-Yong;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
    • /
    • v.35 no.3
    • /
    • pp.215-224
    • /
    • 2000
  • In this study, the distribution of the mec regulator genes and the presence of the mutation in mecI gene and mec promoter region among 50 MRSA clinical isolates derived from a single university hospital in Korea were analyzed. Among 50 MRSA strains, 13 strains had a deletion of mecI gene, and 37 strains were found to have mutations in mecI gene or mecA promoter region corresponding to a presumptive operator of mecA, i.e., the binding site of the repressor protein. Furthermore, in order to track the evolution of methicillin-resistant Staphylococcus aureus (MRSA) distributed in Korea, we determined the MRSA clonotype by combined use of genetic organization patterns of mec regulator genes, ribotype, and coagulase type. As the result, 48 of 50 MRSA strains could be classified into four distinct clones. Clonotype I is characterized by the coagulase type 3, deletion of mecI gene, and ribotype 1 shared by NCTC10442, the first reported MRSA isolate in England (9 strains). Clonotype II is characterized by the coagulase type 4, C to T substitution at position 202 of mecI gene, and ribotypes 2, 3 and 4 shared by 85/3619 strain isolated in Austria (10 strains). Clonotype III is characterized by the coagulase type 2, mutations of mecA promoter region and/or mecI, and ribotypes 4, 5, and 6 shared by N315 strain isolated in Japan (25 strains). Clonotype IV is characterized by the coagulase type 4, deletion of mecI gene, and ribotype 7 (4 strains). The clonality of two strains could not be determined due to their undefined ribotype.

  • PDF