• Title/Summary/Keyword: molecular biological techniques

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Ruminal ciliates as modulators of the rumen microbiome

  • Tansol Park
    • Animal Bioscience
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    • 제37권2_spc호
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    • pp.385-395
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    • 2024
  • Ruminal ciliates are a fundamental constituent within the rumen microbiome of ruminant animals. The complex interactions between ruminal ciliates and other microbial guilds within the rumen ecosystems are of paramount importance for facilitating the digestion and fermentation processes of ingested feed components. This review underscores the significance of ruminal ciliates by exploring their impact on key factors, such as methane production, nitrogen utilization efficiency, feed efficiency, and other animal performance measurements. Various methods are employed in the study of ruminal ciliates including culture techniques and molecular approaches. This review highlights the pressing need for further investigations to discern the distinct roles of various ciliate species, particularly relating to methane mitigation and the enhancement of nitrogen utilization efficiency. The promotion of establishing robust reference databases tailored specifically to ruminal ciliates is encouraged, alongside the utilization of genomics and transcriptomics that can highlight their functional contributions to the rumen microbiome. Collectively, the progressive advancement in knowledge concerning ruminal ciliates and their inherent biological significance will be helpful in the pursuit of optimizing rumen functionality and refining animal production outcomes.

Cellular and Molecular Pathology of Fungi on Plants Studied by Modern Electron Microscopy

  • Sanwald, Sigrun-Hippe
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 1995년도 Proceedings of special lectures on Molecular Biological Approaches to Plant Disease National Agricultural Science and Technology Institute Suwon, Korea
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    • pp.27-53
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    • 1995
  • In plant pathology there is an increasing necessity for improved cytological techniques as basis for the localization of cellular substances within the dynamic fine structure of the host-(plant)-pathogen-interaction. Low temperature (LT) preparation techniques (shock freezing, freeze substitution, LT embedding) are now successfully applied in plant pathology. They are regarded as important tools to stabilize the dynamic plant-pathogen-interaction as it exists under physiological conditions. - The main advantage of LT techniques versus conventional chemical fixation is seen in the maintenance of the hydration shell of molecules and macromolecular structures. This results in an improved fine structural preservation and in a superior retention of the antigenicity of proteins. - A well defined ultrastructure of small, fungal organisms and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. From analyses of the compatible interaction between Erysiphe graminis f. sp. hordei on barley (Hordeum vulgare), various steps in the infection sequence can be identified. Infection sites of powdery mildew on primary leaves of barley are analysed with regard to the fine structural preservation of the haustoria. The presentation s focussed on the ultrastructure of the extrahaustorial matrix and the extrahaustorial membrane. - The integration of improved cellular preservation with a molecular analysis of the infected host cell is achieved by the application of secondary probing techniques, i.e. immunocytochemistry. Recent data on the characterization of freeze substituted powdery mildew and urst infected plant tissue by immunogold methodology are described with special emphasis on the localization of THRGP-like (threonine-hydrxyproline-rich glycoprotein) epitopes. Infection sites of powdery mildew on barley, stem rust as well as leaf rust (Puccinia recondita) on primary leaves of wheat were probed with a polyclonal antiserum to maize THRGP. Cross-reactivity with the anti-THRGP antiserum was observed over the extrahaustorial matrix of the both compatible and incompatible plant-pathogen interactions. The highly localized accumulation of THRGP-like epitopes at the extrahaustorial host-pathogen interface suggests the involvement of structural, interfacial proteins during the infection of monocotyledonous plants by obligate, biotrophic fungi.

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Microbial Structure and Community of RBC Biofilm Removing Nitrate and Phosphorus from Domestic Wastewater

  • Lee, Han-Woong;Choi, Eui-So;Yun, Zu-Whan;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • 제18권8호
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    • pp.1459-1469
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    • 2008
  • Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100${\mu}m$. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was $\beta$- and $\gamma$-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250${\mu}m$. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250${\mu}m$, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250${\mu}m$. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate removal by an RBC system.

환경 오염물질의 진보된 독성 평가 기법 (Recent Advanced Toxicological Methods for Environmental Hazardous Chemicals)

  • 류재천;최윤정;김연정;김형태;방형애;송윤선
    • Environmental Analysis Health and Toxicology
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    • 제14권1_2호
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    • pp.1-12
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    • 1999
  • Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

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향기농업: 휘발성 물질을 이용한 식물병 진단과 방제 (Aromatic Agriculture: Volatile Compound-Based Plant Disease Diagnosis and Crop Protection)

  • 류명주;손진수;오상근;류충민
    • 식물병연구
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    • 제28권1호
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    • pp.1-18
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    • 2022
  • 휘발성물질은 자연에서 어디에나 존재한다. 생태학적으로 식물이나 미생물이 생산하는 휘발성물질은 식물-미생물이나 미생물-미생물간 대화에 중요한 역할을 수행한다. 정족수 인식신호는 세균과 세균 사이의 짧은 거리에서만 영향을 미치지만 휘발성물질은 20 cm 이상의 거리에서 생명체 간 신호전달이 가능하다. 이번 리뷰에서는 휘발성물질을 이용한 식물병진단과 진균, 세균, 바이러스병의 생물적방제의 최신 결과를 소개하였다. 더불어 이러한 휘발성물질을 농업에 적용하기 위한 다양한 기술들도 소개하였다. 휘발성물질의 캡슐화와 서방형 제제화 그리고 바이오나노 융합기술은 기존의 휘발성 물질 적용 한계를 넘게 해 줄 것이다. 종합하면 휘발성물질은 식물병을 효과적으로 방제할 수 있는 새로운 방법이다. 이번 리뷰를 통하여 농민들과 젊은 연구자들이 휘발성물질에 대한 이해를 높이고 향기농업으로의 전환을 앞당기는 계기가 되기를 희망한다.

유전자 발현 데이터를 이용한 암의 유형 분류 기법 (Cancer-Subtype Classification Based on Gene Expression Data)

  • 조지훈;이동권;이민영;이인범
    • 제어로봇시스템학회논문지
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    • 제10권12호
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    • pp.1172-1180
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    • 2004
  • Recently, the gene expression data, product of high-throughput technology, appeared in earnest and the studies related with it (so-called bioinformatics) occupied an important position in the field of biological and medical research. The microarray is a revolutionary technology which enables us to monitor several thousands of genes simultaneously and thus to gain an insight into the phenomena in the human body (e.g. the mechanism of cancer progression) at the molecular level. To obtain useful information from such gene expression measurements, it is essential to analyze the data with appropriate techniques. However the high-dimensionality of the data can bring about some problems such as curse of dimensionality and singularity problem of matrix computation, and hence makes it difficult to apply conventional data analysis methods. Therefore, the development of method which can effectively treat the data becomes a challenging issue in the field of computational biology. This research focuses on the gene selection and classification for cancer subtype discrimination based on gene expression (microarray) data.

Correlation between EGFR Expression and Radiosensitivity in Cervical Adenocarcinoma Cases

  • Hernowo, Bethy S;Suryanti, Sri;Wibisono, Fennisia
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권5호
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    • pp.2535-2537
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    • 2016
  • Background: Most of the cervical cancer patients, including those with cervical adenocarcinomas, come at advanced stage in the developing world so its mortality is high. Radiotherapy is one of the treatment modality for advanced stage cervical adenocarcinomas, but its efficacy depends on several prognostic factors such as the stage, histopathology, presence of organ dysfunction and expression of cellular biology markers mainly involve in cell proliferation such as the epidermal growth factor receptor (EGFR). Some research indicates that activation of EGFR in malignancy (including cervical cancer) correlates with aggressive behavior, a poor prognosis and decreasing sensitivity of radiotherapy. However, the combination between targeted therapies and radiotherapy are innovative approaches which may provide a good result. This study aimed to assess any correlation between expression of EGFR and response to radiotherapy in cervical adenocarcinoma cases. Materials and Methods: A total of 32 women were registered in a retrospective study period January 2007 and May 2014. Paraffin blocks from these patients were processed by classical histological techniques and for immunohistochemical staining of EGFR, scoring being accomplished according to the immunoreactive scoring (IRS) of Remmele and Stegner. Results: Among the studied molecular factors, there was significant correlation expression of EGFR with poor response to radiotherapy (p=0.0001). Conclusions: The result of this study showed a significant correlation between expression of EGFR and sensitivity of radiation in cervical adenocarcinoma cases. Further research is necessary to obtain information about new therapeutic management.

Agrobacterium-mediated Transformation of Rehmannia glutinosa L. with Resveratrol Gene (RS3) of Peanut

  • Lim, Jung-Dae;Yang, Deok-Chun;Yun, Song-Joong;Chung, Ill-Min;Sung, Eun-Soo;Kim, Myong-Jo;Heo, Kweon;Yu, Chang-Yeon
    • 한국약용작물학회지
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    • 제12권2호
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    • pp.171-178
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    • 2004
  • The objectives of this study were to establish the genetic transformation system of stilbene synthase in Rehmannia glutinosa. Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Stilbene synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. PCR analysis with RS3 primer confirmed that the targeted gene was introduced into the plant genome, 904 bp in size. Further analyses of identification of transformation using developed other molecular techniques and transgenic plants that RS t-DNA introduced to chinese foxglove (R. glutinosa L) and its reaction product, stilbene such as resveratrol will be isolate and characterize using NMR, MS, and HPLC.

Local protein synthesis in neuronal axons: why and how we study

  • Kim, Eunjin;Jung, Hosung
    • BMB Reports
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    • 제48권3호
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    • pp.139-146
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    • 2015
  • Adaptive brain function and synaptic plasticity rely on dynamic regulation of local proteome. One way for the neuron to introduce new proteins to the axon terminal is to transport those from the cell body, which had long been thought as the only source of axonal proteins. Another way, which is the topic of this review, is synthesizing proteins on site by local mRNA translation. Recent evidence indicates that the axon stores a reservoir of translationally silent mRNAs and regulates their expression solely by translational control. Different stimuli to axons, such as guidance cues, growth factors, and nerve injury, promote translation of selective mRNAs, a process required for the axon's ability to respond to these cues. One of the critical questions in the field of axonal protein synthesis is how mRNA-specific local translation is regulated by extracellular cues. Here, we review current experimental techniques that can be used to answer this question. Furthermore, we discuss how new technologies can help us understand what biological processes are regulated by axonal protein synthesis in vivo.

Protein Sequence Search based on N-gram Indexing

  • Hwang, Mi-Nyeong;Kim, Jin-Suk
    • Bioinformatics and Biosystems
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    • 제1권1호
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    • pp.46-50
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    • 2006
  • According to the advancement of experimental techniques in molecular biology, genomic and protein sequence databases are increasing in size exponentially, and mean sequence lengths are also increasing. Because the sizes of these databases become larger, it is difficult to search similar sequences in biological databases with significant homologies to a query sequence. In this paper, we present the N-gram indexing method to retrieve similar sequences fast, precisely and comparably. This method regards a protein sequence as a text written in language of 20 amino acid codes, adapts N-gram tokens of fixed-length as its indexing scheme for sequence strings. After such tokens are indexed for all the sequences in the database, sequences can be searched with information retrieval algorithms. Using this new method, we have developed a protein sequence search system named as ProSeS (PROtein Sequence Search). ProSeS is a protein sequence analysis system which provides overall analysis results such as similar sequences with significant homologies, predicted subcellular locations of the query sequence, and major keywords extracted from annotations of similar sequences. We show experimentally that the N-gram indexing approach saves the retrieval time significantly, and that it is as accurate as current popular search tool BLAST.

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