• Title/Summary/Keyword: blast analysis

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MOLECULAR CLONING AND SEQUENCE ANALYSIS OF THE GENE FOR THE HEMIN-BINDING PROTEIN FROM Prevotella intermedia (Prevotella intermedia에서의 Hemin 결합 단백질 유전자의 분리 및 염기서열 분석)

  • Kim, Shin;Kim, Sung-Jo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.304-310
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    • 2006
  • Prevotella intermedia is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. This study has identified a hemin-binding P. intermedia protein by expression of a P. intermedia genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. The genomic library of P. intermedia was constructed into plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$, and screened for recombinant clones using heminbinding activity by plating onto hemin-containing agar. Approximately 5,000 recombinant E. coli colonies were screened onto LB-amp-hemin agar, single clone(pHem1) was exhibited a clearly pigmented phonotype. The 2.5kb insert DNA of pHem1 was determined by restriction enzyme mapping. Southern blot analysis of BamHI, BglII, EcoRI, HindIII and PstI-digested P. intermedia DNA indicated that single copy of the gene was present in the genome. Northern blot analysis revealed that the size of transcript was approximately 1.8 kb. The cloned gene contained a single ORF, consisting of approximately 850-residue amino acids. A BLAST search of the Institute for Genomic Research genes with similar nucleotide sequence revealed no significant similarity It needs further investigation to clarify the mechanisms of heme uptake in P. intermedia.

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Genetic Species Identification by Sequencing Analysis of Nuclear and Mitochondrial Genes for Albino Misgurnus Species from Korea (우리나라 미꾸리속(genus Misgurnus) 알비노 개체의 미토콘드리아 및 핵 유전자 염기서열 분석에 의한 유전적 동정)

  • Song, Ha-Youn;Moon, Shin-Joo;Kim, Keun-Sik;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.29 no.2
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    • pp.139-145
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    • 2017
  • The spontaneous color mutant, albino individuals of genus Misgurnus, are rarely discovered in Korea and there are difficult to identify morphological species due to lack melanin pigmentation. In this study, we developed a genetic identification method for the species of albino Misgurnus individuals based on phylogenetic analysis by using recombination activating gene 1 (rag1) and cytochrome b (cytb) region of mitochondrial DNA. As a result of molecular phylogenetic analysis, three clades were identified as Misgurnus mizolepis, M. anguillicaudatus and M. mohoity. The homology of the cytb sequences of M. mohoity was best match to that of M. mohoity sequences in GenBank database. As a result of species identification of 25 albino Misgurnus individuals based on the phylogenetic tree, the red-eye type was identified as 16 M. anguillicaudatus and one M. mizolepis. The remaining three individuals were identified as one M. mizolepis ♀${\times}$M. anguillicaudatus ♂, and two M. mohoity ♀${\times}$M. anguillicaudatus ♂, respectively. In addition, the five black-eye type individuals were identified as one M. anguillicaudatus, three M. mizolepis and one M. mohoity. Therefore, this genetic identification method will be an useful techniques for species or hybrid identification in genus Misgurnus.

Multilateral analysis of $Renova^{(R)}$ implant placement and its Survival rate ($Renova^{(R)}$ 임플란트 식립 후 단기간의 생존율에 대한 다각적 분석)

  • Yang, Jin-Hyuk;Kim, Sung-Tae;Jung, Ui-Won;Nam, Woong;Jung, Young-Soo;Shim, June-Sung;Moon, Hong-Seok;Lee, Keun-Woo;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.38 no.3
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    • pp.413-428
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    • 2008
  • Purpose: Given the predictability of dental implant procedure from the studies of successful osseointegration, implant dentistry is often the treatment of choice to replace missing teeth in edentulous patient instead of the fixed prosthesis or removable denture. The $Renova^{(R)}$ dental implant has a RBM(Resorbable Blast Media) surface, internal hex prosthetic connection and a tapered design. At this study gives the analysis of the implant and the short term survival rate of the implant. Material and Methods: In this study, a multilateral analysis was performed on the subjects undergoing placement with $Renova^{(R)}$ implant between August 2006 and February 2008 in Yonsei University dental hospital. 96 implants were placed in 56 patients and they were surveyed for cumulative survival rate. Among them 78 implants in 44 patients were surveyed for the rest analyses. Result: 1. The cumulative survival rate was 96.88% of 96 implants in 56 patients. 2. The mean marginal bone loss was 0.803mm and the marginal bone loss in augmentation group has higher value than the marginal bone loss in non augmentation group. 3. The health scale for the implants were 87% in success group, 9% in satisfactory survival group, 1% in compromised survival group, and 3% in failure group. 4. Two implants placed in poor bone posterior area by 2-stage failed during prosthetic procedure. Conclusion: $Renova^{(R)}$ dental implant showed high cumulative survival rate in installation on partial edentulous ridge and could be a predictable implant system.

Flexural Analysis of RC Beam Considering Autogenous Shrinkage Model (자기수축 모델을 고려한 철근콘크리트 보의 휨 거동 해석)

  • Yoo Sung-Won;Soh Yang-Sub;Cho Min-Jung;Koh Kyung-Taek;Jung Sang-Hwa
    • Journal of the Korea Concrete Institute
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    • v.17 no.4 s.88
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    • pp.621-628
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    • 2005
  • Recently, it is noticed that autogenous shrinkage of high-performance concrete causes early crack in high performance concrete structures. The purpose of the present study is to derive a realistic equation to estimate the autogenous shrinkage of high performance concrete and to apply to structural analysis. For this purpose, several series of concrete specimens have been tested. When water-binder ratio is fixed to $30\%$, major test variables were the type and contents of mineral admixture. The autogenous shrinkage of HPC with fly ash slightly decreased than that of OPC concrete, but the use of blast furnace slag increased with the autogenous shrinkage. A prediction equation to estimate the autogenous shrinkage of HPC with mineral admixture was derived and proposed in this study. The proposed equation show reasonably good correlation with test data on autogenous shrinkage of HPC with mineral admixture. The finite element program developed in this study provides the useful tool for the flexural analysis including the autogenous shrinkage model. By this program, we know that the tensile stress considering the autogenous shrinkage of reinforced concrete structures increase $20\~27\%$ than that not considering.

Development of the HEMP Generation, Propagation Analysis, and Optimal Shelter Design Tool (고 고도 전자기파(HEMP) 발생과 전파해석 및 방호실 최적 설계 Tool 개발)

  • Kim, Dong Il;Min, Gyeong Chan
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.10
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    • pp.2331-2338
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    • 2014
  • The HEMP threat may have acquired new, and urgent, relevance as the proliferation of nuclear weapons and missile technology accelerates of the North Korea, for example, is assessed as already having developed few atomic weapons, and is on the verge of North Korea already has missiles capable of delivering a nuclear warhead against South Korea. ITU K.78, K81 and IEC recommended its counter-measuring for the industrial facilities with navigation and sailing facilities in order to obviate the all of processor equipped system malfunctions from the EMP/HEMP but its simulation must only be done by the computer simulation which had studied on the 1960-1990 years USA/AFWL papers. This result has a significant activities to the South Korea being under the North Korea threat because all of HEMP related products was strongly limited for export. The HEMP cord which was developed newly by the KTI including the HEMP generation & propagation analysis, optimal shelter design tool, essential EM energy attenuation in multi-layered various soils and rocks and HEMP filter design tool. Especially, the least square fitting method was adopted to analysis for the EM energy attenuation in the soils and rocks because it has a various characteristics based on the many times field test reports.

Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae (동양달팽이(Nesiohelix samarangae)의 arginine kinase 유전자 분석 및 발현 패턴에 관한 연구)

  • Jeong, Ji Eun;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.171-179
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    • 2013
  • Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

  • Meilina, Lita;Budiarti, Sri;Mustopa, Apon Zaenal;Darusman, Huda Shalahudin;Triratna, Lita;Nugraha, Muhammad Ajietuta;Bilhaq, Muhammad Sabiq;Ningrum, Ratih Asmana
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.75-87
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    • 2021
  • Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPUsp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC50 values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Effects of Magnesium and Sulfate Ions on the Sulfate Attack Resistance of Alkali-activated Materials (알칼리 활성화 결합재 모르타르의 황산염 침식 저항성에 미치는 마그네슘 및 황산 이온의 영향)

  • Park, Kwang-Min;Cho, Young-Keun;Shin, Dong-Cheol
    • Journal of the Korea Concrete Institute
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    • v.29 no.4
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    • pp.415-424
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    • 2017
  • The purpose of this study is to investigate the effect of sulfate (${SO_4}^{2-}$) and magnesium ($Mg^{2+}$) ions on sulfate resistance of Alkali-activated materials using Fly ash and Ground granulated blast furnace slag (GGBFS). In this research, 30%, 50% and 100% of GGBFS was replaced by sodium silicate modules ($Ms(SiO_2/Na_2O)$, molar ratio, 1.0, 1.5 and 2.0). In order to investigate the effects of $Mg^{2+}$ and ${SO_4}^{2-}$, compression strength, weight change, lengh expansion of the samples were measured in 10% sodium sulfate ($Na_2SO_4$), 10%, 5% and 2.5% magnesium sulfate ($MgSO_4$), 10% magnesium nitrate ($Mg(NO_3)_2$), 10% [magnesium chloride ($MgCl_2$) + sodium sulfate ($Na_2SO_4$)] and 10% [magnesium nitrate $(Mg(NO_3)_2$ + sodium sulfate ($Na_2SO_4$)] solution, respectively and X-ray diffraction analysis was conducted after each experiment. As a result, when $Mg^{2+}$ and ${SO_4}^{2-}$ coexist, degradation of compressive strength and expansion of the sample were caused by sulfate erosion. It was found that the reaction of $Mg^{2+}$ with Calcium Silicate Hydrate (C-S-H) occurred and $Ca^{2+}$ was produced. Then the Gypsum ($CaSO_4{\cdot}2H_2O$) was formed due to reaction between $Ca^{2+}$ and ${SO_4}^{2-}$, and also Magnesium hydroxide ($Mg(OH)_2$, Brucite) was produced by the reaction between $Mg^{2+}$ and $OH^-$.

Major character analysis of CAX 1 (cation exchanger 1) transgenic rice plants in In Vivo (CAX 1 형질전환체 벼의 In Vivo에서 주요특성 분석)

  • Kim, Kyung-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.375-383
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    • 2009
  • This study was carried out to develop transgenic rice cultivars with the CAX1 (accession no. U57411) gene. We successfully selected the transgenic rice plants over-expressing the Arabidopsis H+/$Ca^{2+}$ antiporter CAX1 (accession no. U57411) gene in T6 generation. The brown rice of the CAX1 expressing rice contained 13.4~68.0 % more calcium $(Ca^{2+})$ than that of the wild type and 5 lines were selected based on the phenotypes compared to the control cultivar at the GMO field. The CAX1 expressing transgenic rice plants were similar in phenotype to the wild type during the whole growth period. Also these selected 4 lines appeared to be resistant to blast, cold and water solution compared with the wild type. Difference in 1,000 grain weight of brown rice was observed among each line but grain shape did not show any morphological alternations. These results suggest the enhanced Ca-substrate specificity of CAX1 exchanger in donor plant. Therefore, intact CAX1 exchanger can be functionally useful for $Ca^{2+}$ nutrient enrichment of rice with reduced accumulation of undesirable cation.

Identification of a New Potyvirus, Keunjorong mosaic virus in Cynanchum wilfordii and C. auriculatum (큰조롱과 넓은잎 큰조롱에서 신종 포티바이러스(큰조롱모자이크바이러스)의 동정)

  • Lee, Joo-Hee;Park, Seok-Jin;Nam, Moon;Kim, Min-Ja;Lee, Jae-Bong;Sohn, Hyoung-Rac;Choi, Hong-Soo;Kim, Jeong-Soo;Lee, Jun-Seong;Moon, Jae-Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.238-246
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    • 2010
  • In 2006 fall, a preliminary survey of viruses in two important medicinal plants, Cynanchum wilfordii and C. auriculatum, was conducted on the experimental fields at the Agricultural Research and Extension Services of Chungbuk province in Korea. On each experimental fields, percentage of virus infection was ranged from 20 to 80%, and especially an average of disease incidence propagated by roots was twice higher than that by seeds. The various symptoms were observed in Cynanchum spp. plants, such as mosaic, mottle, necrosis, yellowing, chlorotic spot and malformation etc. In electron microscopic examination of crude sap extracts, filamentous rod particles with 390-730 nm were observed in most samples. The virus particles were purified from the leaves of C. wilfordii with typical mosaic symptom, and the viral RNA was extracted from this sample containing 430-845 nm long filamentous rod. To identify the viruses, reverse transcription followed by PCR with random primers was carried out. The putative sequences of P3 and coat protein of potyvirus were obtained. From a BLAST of the two sequences, they showed 26-38% and 62-72% identities to potyviruses, respectively. In SDS-PAGE analysis, the subunit of coat protein was approximately 30.3 kDa, close to the coat protein of potyvirus. In bioassay with 21 species in 7 families, Chenopodium quinoa showed local lesion on inoculated leave and chlorotic spot on upper leave, but the others were not infected. RT-PCR detection using specific primer of C. wilfordii and C. auriculatum samples, all of 24 samples with virus symptom was positive, and five out of seven samples without virus symptom were also positive. On the basis of these data, the virus could be considered as a new member of potyvirus. We suggested that the name of the virus was Keunjorong mosaic virus (KjMV) after the common Korean name of C. wilfordii.