• 제목/요약/키워드: molecular biological techniques

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과수육종에 있어 생명공학의 이용 전망 (The Prospectss and Utilization of Biotechnology for the Improvement of Fruit Breeding)

  • 이돈균;김휘천;신용억;강상조;예병우
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1995년도 제9회 식물생명공학 심포지움 식물육종과 분자생물학의 만남 The 9th Plant Biotechnology Symposium -Breeding and Molecular Biology-
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    • pp.133-170
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    • 1995
  • The major objectives of fruit breeding lie in improvement of cultivar, easy to be cultivated and of high quality, in order to produce unexpensive, delicious fruit both for fresh fruit market and processing. Recently, fruit breeding in Korea has contributed to breeding of several superior cultivars in major fruit crops, resulting in appreciable improvement in qualities such as skin color, taste and fruit-bearing habit concerned with productivity. In spite of accomplishments mentioned above, the need for both highly disease-resistant cultivars and long-keeping, physiological disorder-resistant cultivars to meet long distance transsportation in the temperate fruit crops of apples, oriental pears, stone fruits such as peaches, and grapes grown in Korea is rapidly pressing more than ever, as cultivars of today susceptible to pests and diseases and vulnerable to physiological disorders are very expensive and time-consuming in post-harvest handling and management. Thus, imporvements made in the above problems through breeding level will lead to the really enhanced productivity in fruit industry. The major impediments of tree size, the long length of juvenile period and the highly heterogeneous genetic composition to the improvement of fruit crops are responsible for the lower amount and rate of improvements of fruit crops as compared to annuals. Considering the expected limitations of the above problems to be solved through conventional breeding methods and strategy, a turning point of breeding a near perfect cultivar would be laid down if innovative breakthroughs in biological technology will be realized in applying some of the techniques of genetic manipulation at the molecular level to the cultivar improvement of fruit crops, such as the selective insertion of DNA carrying genes that govern desirable characteristics. More than anything else, those traits such as fruiting habit deciding productivity, elements of fruit qualities conditioned by monogene, and disease-and pest-resistance of vital importance for successful fruit growing are urgently desired to be improved by advancement of biotechnology for they are more than difficult and need long period to be attained through conventional breeding method.

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대규모 유전자 분석 기법을 이용한 육미지황원의 유전자 발현 연구 (Studies on Gene Expression of Yukmijihwang-tang using High-throughput Gene Expression Analysis Techniques)

  • 강봉주;김윤택;조동욱
    • 한국한의학연구원논문집
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    • 제8권2호통권9호
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    • pp.95-107
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    • 2002
  • Yukmijihwang-tang(YM) is a noted herbal prescription in Chinese and Korean traditional medicines, and it has been known to reinforce the vital essence and has been widely used for a variety of disease such as stroke, osteoporosis, anti-tumor, and hypothyrodism. Regarding its traditional use, YM has been known to reinforce the Yin (vital essence) of liver and kidney. Also it has been known to reinforce nutrition and biological function in brain. Recently, studies suggested that YM increase antioxidant activities and exert the protective effect against oxidant-induced liver cell injury. We investigated the high-throughput gene expression analysis on the Yukmijihwang-tang administrated in SD rats. Microarray data were validated on a limited number of genes by semiquantitative RT-PCR and Western blot analyses. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes in drug discovery This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with herbal medicine. Total RNA from normal control brain and Yukmijihwang-tang administrated brain were hybridized to microarrays containing 10,000 rat genes. The 52 genes were found to be up-regulated(twice or more) excluding EST gene. The nine genes were found to be down-regulated(twice or more) excluding EST gene. Gene array technology was used to identify for the first time many genes expression pathway analysis that arecell cycle pathway, apoptosis pathway, electron transport chain pathway, cytoplasmic ribosomal protein pathway, fatty acid degradation pathway, and TGF-beta signaling pathway. These differentially expressed genes pathway analysis have not previously been iavestigated in the context of herbal medicine efficacy and represent novel factors for further study of the mechanism of herbal medicine efficacy.

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한국재래산양의 핵형분석 (The karyotype of Korean native goat (capra hircus))

  • 오승현;윤영민;윤여성;이준섭;이흥식;성제경
    • 대한수의학회지
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    • 제39권5호
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    • pp.908-920
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    • 1999
  • We investigated the cytogenetic characteristics of Korean native goat(Capra hircus). Chromosome slides were prepared from peripheral blood cell cultures. GTG, GBG, RBG and CBG-banding techniques were employed on those slides. The high resolution karyotype of Korean native goat could be made with the incorporation of BrdU. Korean native goat has 60 chromosomes composed of 58 autosomes and XY or XX sex chromosomes. All of autosomes of Korean native goat were acrocentric chromosomes. X chromosome was submetacentric and Y chromosome was metacentric. The GTG, GBG and RBG-band patterns of Korean native goat were similar to those of other goats. CBG-band regions were distinct at the proximal portion of the long arms of all autosomes in Korean native goats. According to our investigation, there was no significant difference in chromosomal band patterns between Korean native goat and other goats. It might be necessary to use molecular genetic markers for clarifying the genetical characteristics of Korean native goat whose biological characteristics are not clearly defined.

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분자 세포 유전학 기법에 응용되는 영상 처리 기술 (Image Analysis Algorithms for Comparative Genomic Hybridization)

  • 김대석;유진성;이진우;김종원;문신용;최영민
    • 대한의용생체공학회:학술대회논문집
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    • 대한의용생체공학회 1998년도 추계학술대회
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    • pp.66-69
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    • 1998
  • Comparative Genomic Hybridization (CGH)은 세포 내 특정 DNA 서열 이상을 염색체상에 보여주는 중요한 분자 세포 유전학 기법이다. CGH 기법에서는 세포 분열 중기의 염색체에서 준비한 형광 비율 영상의 정량적 분석을 위해서 Digital 영상 처리 기술이 쓰여야 한다. 본 논문에서는 최근 연구 개발된 영상 처리 algorithm들이 어떻게 CGH 기법에 쓰이는 지를 소개하려 한다. 각 염색체의 형광 비율 profile를 평균하기 위해, 염색체 영상의 이원화, 염색체 영상 뼈대 변환(skeletonization), 뼈대 정보의 변수화와 영상 명암의 재추출을 통한 굽은 염색체 영상 펴기 등이 언구되었다. 개발된 algorithm 들은 바이오메드랩 사의 ChIPS 핵형 정렬 시스템에 구현했다.

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나노입자의 현황조사 및 처리방안 마련을 위한 문헌연구 (Review of Nanoparticles in Drinking Water: Risk Assessment and Treatment)

  • 김승현;홍승관;윤제용;김두일;이상호;권지향;김형수;독고석;국지훈
    • 상하수도학회지
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    • 제25권2호
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    • pp.201-212
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    • 2011
  • Nanotechnology is the applied science which develops new materials and systems sized within 1 to 100 nanometer, and improves their physical, chemical, and biological characteristics by manipulating on an atomic and molecular scale. This nanotechnology has been applied to wide spectrum of industries resulting in production of various nanoparticles. It is expected that more nanoparticles will be generated and enter to natural water bodies, imposing great threat to potable water resources. However their toxicity and treatment options have not been throughly investigated, despite the significant growth of nanotechnology-based industries. The objective of this study is to provide fundamental information for the management of nanoparticles in water supply systems through extensive literature survey. More specifically, two types of nanoparticles are selected to be a potential problem for drinking water treatment. They are carbon nanoparticles such as carbon nanotube and fullerene, and metal nanoparticles including silver, gold, silica and titanium oxide. In this study, basic characteristics and toxicity of these nanoparticles were first investigated systematically. Their monitoring techniques and treatment efficiencies in conventional water treatment plants were also studied to examine our capability to mitigate the risk associated with nanoparticles. This study suggests that the technologies monitoring nanopartilces need to be greatly improved in water supply systems, and more advanced water treatment processes should be adopted for better control of these nanoparticles.

Analysis of Microbial Communities Using Culture-dependent and Culture-independent Approaches in an Anaerobic/Aerobic SBR Reactor

  • Lu Shipeng;Park Min-Jeong;Ro Hyeon-Su;Lee Dae-Sung;Park Woo-Jun;Jeon Che-Ok
    • Journal of Microbiology
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    • 제44권2호
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    • pp.155-161
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    • 2006
  • Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

Structural Bioinformatics Analysis of Disease-related Mutations

  • Park, Seong-Jin;Oh, Sang-Ho;Park, Dae-Ui;Bhak, Jong
    • Genomics & Informatics
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    • 제6권3호
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    • pp.142-146
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    • 2008
  • In order to understand the protein functions that are related to disease, it is important to detect the correlation between amino acid mutations and disease. Many mutation studies about disease-related proteins have been carried out through molecular biology techniques, such as vector design, protein engineering, and protein crystallization. However, experimental protein mutation studies are time-consuming, be it in vivo or in vitro. We therefore performed a bioinformatic analysis of known disease-related mutations and their protein structure changes in order to analyze the correlation between mutation and disease. For this study, we selected 111 diseases that were related to 175 proteins from the PDB database and 710 mutations that were found in the protein structures. The mutations were acquired from the Human Gene Mutation Database (HGMD). We selected point mutations, excluding only insertions or deletions, for detecting structural changes. To detect a structural change by mutation, we analyzed not only the structural properties (distance of pocket and mutation, pocket size, surface size, and stability), but also the physico-chemical properties (weight, instability, isoelectric point (IEP), and GRAVY score) for the 710 mutations. We detected that the distance between the pocket and disease-related mutation lay within $20\;{\AA}$ (98.5%, 700 proteins). We found that there was no significant correlation between structural stability and disease-causing mutations or between hydrophobicity changes and critical mutations. For large-scale mutational analysis of disease-causing mutations, our bioinformatics approach, using 710 structural mutations, called "Structural Mutatomics," can help researchers to detect disease-specific mutations and to understand the biological functions of disease-related proteins.

A novel pattern recognition protein of the Chinese oak silkmoth, Antheraea pernyi, is involved in the pro-PO activating system

  • Wang, Xialu;Zhang, Jinghai;Chen, Ying;Ma, Youlei;Zou, Wenjun;Ding, Guoyuan;Li, Wei;Zhao, Mingyi;Wu, Chunfu;Zhang, Rong
    • BMB Reports
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    • 제46권7호
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    • pp.358-363
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    • 2013
  • In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti-Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein.

주요 오염물질로 오염된 지하수에서 미생물의 무배양식 군집분석방법과 미생물상에 대한 조사방법 연구 (Culture-Independent Methods of Microbial Community Structure Analysis and Microbial Diversity in Contaminated Groundwater with Major Pollutants)

  • 김재수
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제11권3호
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    • pp.66-77
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    • 2006
  • 최근에 적용된 지하수 미생물의 군집구조를 밝히는 분자생물학적 및 생화학적 방법들에 대해서 알아보았고 그 결과로서 지하수의 주요 오염물질에 따른 활성화된 미생물군집들이 무엇인지를 밝힌 연구논문들을 종합하여 정리하였다. PCR에 의한 유전자 증폭기술의 발달로 배양 없이 미생물 종류와 개체군을 파악할 수 있게 되었고 각종 finger-printing 방법 (DGGE, SSCP, RISA, microarray) 과 지방산분석법 (PLFA/FAME)을 이용하여 활성화 된 미생물군집구조를 분석하였으며 FISH 등의 방법으로 특정균의 활성도를 알아본 사례들을 조사하였다. 대표적인 지하수오염물질인 유류성분 (n-alkanes, BTEX, MTBE, ethanol)과 염소계 용매 (TCE, PCE, PCB, CE, carbon tetrachloride, chloro-benzene) 등으로 오염되었을 때 우점하는 지하수 미생물상에 대해 보고된 내용을 포함하였다.

Methanol이 배양된 흰쥐 해마의 신경세포 및 신경교 세포의 성장에 미치는 영향 (Effect of Methanol on Cultured Neuronal and Glial Cells on Rat Hippocampus)

  • 이정임;조병채;배영숙;이경은
    • Toxicological Research
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    • 제12권2호
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    • pp.203-211
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    • 1996
  • Methanol has been widely used as an industrial solvent and environmental exposure to methanol would be expected to be increasing. In humans, methanol causes metabolic acidosis and damage to ocular system, and can lead to death in severe and untreated case. Clinical symptoms are attributed to accumulation of forrnic acid which is a metabolic product of methanol. In humans and primates, formic acid is accumulated after methanol intake but not in rodents due to the rapid metabolism of methanol. Neverthless, the developmental and reproductive toxicity were reported in rodents. Previous reports showed that perinatal exposure to ethanol produces a variety of damage in human central nervous system by direct neurotoxicity. This suggests that the mechanism of toxic symptoms by methanol in rodents might mimic that of ethanol in human. In the present study I hypothesized that methanol can also induce toxicity in neuronal cells. For the study, primary culture of rat hippocampal neurons and glias were empolyed. Hippocampal cells were prepared from the embryonic day-17 fetuses and maintained up to 7 days. Effect of methanol (10, 100, 500 and 1000 mM) on neurite outgrowth and cell viability was investigated at 0, 18 and 24 hours following methanol treatment. To study the changes in proliferation of glial cells, protein content was measured at 7 days. Neuronal cell viability in culture was not altered during 0-24 hours after methanol treatment. 10 and 100 mM methanol treatment significantly enhanced neurite outgrowth between 18-24 hours. 7-day exposure to 10 or 100 mM methanol significantly increased protein contents but that to 1000 mM methanol decreased in culture. In conclusion, methanol may have a variety of effects on growing and differentiation of neurons and glial cells in hippocampus. Treatment with low concentration of methanol caused that neurite outgrowth was enhanced during 18-24 hours and the numbers of glial cell were increased for 7 days. High concentration of methanol brought about decreased protein contents. At present, the mechanism responsible for the methanol- induced enhancement of neurite outgrowth is not clear. Further studies are required to delineate the mechanism possibly by employing molecular biological techniques.

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