• Title/Summary/Keyword: probe level analysis

Search Result 94, Processing Time 0.025 seconds

$V_H$ Gene Expression and its Regulation on Several Different B Cell Population by using in situ Hybridization technique

  • Jeong, Hyun-Do
    • Journal of fish pathology
    • /
    • v.6 no.2
    • /
    • pp.111-122
    • /
    • 1993
  • The mechanism by which $V_H$ region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of $V_H$ genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed $V_H$ gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed $V_H$ gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive $C{\mu}$ and $V_{H}J558$ RNA probes are tested on LPS-stimulated adult spleen cells, $2{\sim}4{\times}106cpm$/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at $54^{\circ}C$ is found to be optimum for the $C{\mu}$. $V_{H}S107$ and $V_{H}J558$ probes. The analyzed results clearly demonstrate that the level of each different $V_H$ gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of $V_H$ gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific $V_H$ gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.

  • PDF

Correlation analysis of periodontal tissue dimensions in the esthetic zone using a non-invasive digital method

  • Kim, Yun-Jeong;Park, Ji-Man;Cho, Hyun-Jae;Ku, Young
    • Journal of Periodontal and Implant Science
    • /
    • v.51 no.2
    • /
    • pp.88-99
    • /
    • 2021
  • Purpose: Direct intraoral scanning and superimposing methods have recently been applied to measure the dimensions of periodontal tissues. The aim of this study was to analyze various correlations between labial gingival thickness and underlying alveolar bone thickness, as well as clinical parameters among 3 tooth types (central incisors, lateral incisors, and canines) using a digital method. Methods: In 20 periodontally healthy subjects, cone-beam computed tomography images and intraoral scanned files were obtained. Measurements of labial alveolar bone and gingival thickness at the central incisors, lateral incisors, and canines were performed at points 0-5 mm from the alveolar crest on the superimposed images. Clinical parameters including the crown width/crown length ratio, keratinized gingival width, gingival scallop, and transparency of the periodontal probe through the gingival sulcus were examined. Results: Gingival thickness at the alveolar crest level was positively correlated with the thickness of the alveolar bone plate (P<0.05). The central incisors revealed a strong correlation between labial alveolar bone thickness at 1 and 2 mm, respectively, inferior to the alveolar crest and the thickness of the gingiva at the alveolar crest line (G0), whereas G0 and labial bone thickness at every level were positively correlated in the lateral incisors and canines. No significant correlations were found between clinical parameters and hard or soft tissue thickness. Conclusions: Gingival thickness at the alveolar crest level revealed a positive correlation with labial alveolar bone thickness, although this correlation at identical depth levels was not significant. Gingival thickness, at or under the alveolar crest level, was not associated with the clinical parameters of the gingival features, such as the crown form, gingival scallop, or keratinized gingival width.

Nature of a Firm, Degree of Cluster Linkages, and Innovation: A Study of Bengaluru High-tech Manufacturing Cluster

  • Chandrashekar, Deepak
    • Asian Journal of Innovation and Policy
    • /
    • v.7 no.1
    • /
    • pp.103-130
    • /
    • 2018
  • It is generally understood that clusters are the promoters of innovation and therefore, the attention of researchers has been increasingly to discern the factors driving innovation among the firms in a cluster, especially in a high-tech cluster. In this study, we identify the variables capturing the nature of a firm that possibly impact the absorptive capacity of a firm and subsequently ascertain their impact on the degree of interactions between a firm, and other firms and associated institutions within and outside a cluster, respectively. Furthermore, we probe the influence of these interactions as a whole on firm-level innovation. The study was carried out in the context of Bengaluru, which houses the densely interconnected network of innovation-intensive high-tech manufacturing firms forming a high-tech manufacturing cluster. Data were drawn from 101 high-tech manufacturing firms belonging to electronics, machine tools, electrical and pharmaceutical industries. Based on the cluster analysis and subsequent graphical analysis on each of the three profiled clusters, it was found that size and origin of a firm have significant impact on the degree of firm's interactions. In turn, higher dynamism of firms in terms of degree of interactions led to higher innovation performance.

Cloning, DNA Sequence Determination, and Analysis of Growth-Associated Expression of the sodF Gene Coding for Fe- and Zn-Containing Superoxide Dismutase of Streptomyces griseus

  • Kim, Ju-Sim;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
    • /
    • v.10 no.5
    • /
    • pp.700-706
    • /
    • 2000
  • Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplamic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acid of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acid (22,430 Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110 bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.

  • PDF

Cloning of a Chitinase Gene of Xanthomonas sp. Isolated from Soil and its Expression in E. coli. (토양에서 분리된 Xanthomonas sp.의 Chitinase 유전자 cloning과 E.coli에서의 발현)

  • Kim, Ho-Sang;Seong, Ki-Young;Eun, Moo-Young;Hwang, Cher-Won
    • Applied Biological Chemistry
    • /
    • v.41 no.2
    • /
    • pp.125-129
    • /
    • 1998
  • Xanthomonas sp. isolated from soil exhibited cell wall lytic activity of Candida albicans and secreted chitinase in chitin media. Especially, the chitinase activity was induced by chitin and reached a maximum level at 3 days culture in chitin media. We constructed genomic library of Xanthomonas sp. using cosmid vector in E. coli. Oligonucleotide probe was synthesized from the consensus sequence corresponding to chitinase active site, which was derived from the comparison of amino acid sequences of bacterial chitinase genes. Using this oligonucleotide probe, we screened the genomic library. By restriction enzyme mapping of the positive clones, we identified 4 independent clones which may contain the chitinase gene. One of the clones, named pXCH1 (1.2 kb insert), was further analyzed. Northern blot analysis indicated that is transcripts, 1 kb and 0.8 kb, were induced by chitin. When the cloned gene was induced by IPTG in E.coli cell, chitinase activity which was secreted onto culture media was not observed. However, when the cell was disrupted by using sonicator and then centrifuged, the supernatant exhibited chitinase activity. SDS-PAGE of the supernatant indicated that about 35 kDa protein was induced by IPTG. From these results, it was concluded that the cloned DNA was one of the chitinase genes of Xanthomonas sp.

  • PDF

Expression Patterns of Bacillus subtilis Diacylglycerol Kinase Gene Induced by Physiological Stimuli (Bacillus subtilis dgk (diacylglycerol kinase) 유전자의 생리적 자극에 의한 유도발현)

  • Lee, Mi-Young;Suh, Seok-Jong;Lee, Jin-Hyung;Song, Bang-Ho;Kim, Jong-Cuk
    • Microbiology and Biotechnology Letters
    • /
    • v.30 no.1
    • /
    • pp.15-20
    • /
    • 2002
  • Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid and it may play a role in signal transduction in Escherichia coli as well as in eukaryotic cells. In addition, DGK is important for microorganisms to adapt to several physiological stimuli. In Bacillus subtilis, the effect of stress on dgk transcription was examined by northern hybridization. The high level of dgk transcription was induced against high osmolarity, low pH value and low temperature. Transcriptional analysis revealed that the dgk gene and dgk upstream locus (ORF2, ORF3 and ORF4) were transcribed as a polycistronic mRNA to form an approximately 2.5 kb transcript.

Performance Analysis of Matching Layer for Level Measurement in CNG (CNG 레벨계측을 위한 Matching layer의 성능분석)

  • Lee, Hongyoon;Kim, Kwonse;Choi, Dooseuk
    • Journal of the Korea Convergence Society
    • /
    • v.9 no.4
    • /
    • pp.143-150
    • /
    • 2018
  • This work conducted the experimental analysis on ultrasonic acoustic matching in order to measure the accuracy of gaseous fuel level values. The experimental devices were used as 12V-DC supply, control T1 board, oscilloscope (DS01072B), ultrasonic probe and pattern table. The research models were designed by ceramic assay which can determine the transmitting and receiving energies. The result of the ringing area could verify the increased characteristics in the order of D (0.180m) < E (0.184m) < B (0.204m) < A (0.234m) < F (0.244m) < C (0.247m) models based on initial 2.9V of the maximum peak voltage. From the experimental results, the model designed by Ø21*3+2t of the matching layer was notable in that the most outstanding directivity energy could be created.

Factors Affecting Introduction of rolC Gene in Lycium chinense Mill. (구기자나무(Lycium chinense Mill.)로의 rolC유전자 도입에 미치는 요인)

  • 박용구;최명석;김병원;정원일;노광수
    • Korean Journal of Plant Tissue Culture
    • /
    • v.22 no.6
    • /
    • pp.329-334
    • /
    • 1995
  • Transformation system of rolC gene, dwarf gene in Lycium chinenese Mill. established by using system. Pin-punctured leaves induced numerous adventious buds in abaxial side when cultured on 3/2 MS medium containing 2.0 mg/L zeatin. Survival rate and shoot regeneration frequency of leaf explants decreased as kanamycin sulfate level increased. Shoot buds were not regenerated on 3/2 MS medium containing 10 mg/L kanamycin sulfate and 2.0 mg/L zeaein. Of the level tested, 10 mg/L of kanamycin sulfate was optimum in selection of kanamycin sulfate resistant plant. Co-culture time of bacteria and leaf explants was affected at the frequency of shoot regeneration and survival of leaf explants. Leaf explants co-cultivated during above 48hr severely decreased survival rate and shooting rate. Best result on survival rate and shooting rate were obtained when exposed for 24 h. 80 explants of 105 leaf explants survived on 3/2 MS medium containing 2.0 mg/L zeatin and 10 mg/L kanamycin sulfate, and 15 shoots was regenerated on the same medium. To select kanamycin sulfate resistant plant, regenerate as cultured on 3/2 MS medium containing 10 mg/L kanamycin sulfate, and obtained 5 kanamycin resistant plants. Southern blot analysis conformed that the rolC gene was incorporated into the genomic DNA of kanamycin resistant plants.

  • PDF

Isolation and Characterization of Eukaryotic Translation Initiation Factor 5A (eIF-5A) from Potato (감자로부터 Eukaryotic Translation Initiation Factor 5A (elF-5A) 유전자의 동정 및 발현 분석)

  • 인준교;신동호;최관삼;양덕춘
    • Korean Journal of Plant Tissue Culture
    • /
    • v.28 no.5
    • /
    • pp.283-287
    • /
    • 2001
  • Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.

  • PDF

A study on the Conducted Noise Reduction in Random PWM (Random PWM 기법을 이용한 전도노이즈 저감)

  • Jeong, Dong-Hyo
    • Proceedings of the KIEE Conference
    • /
    • 2006.10b
    • /
    • pp.154-158
    • /
    • 2006
  • The switching-mode power converter has been widely used because of its features of high efficiency and small weight and size. These features are brought by the ON-OFF operation of semiconductor switching devices. However, this switching operation causes the surge and EMI(Electromagnetic Interference) which deteriorate the reliability of the converter themselves and entire electronic systems. This problem on the surge and noise is one of the most serious difficulties in AC-to-DC converter. Random Pulse Width Modulation (RPWM) is peformed by adding a random perturbation to switching instant while output-voltage regulation of converter is performed. RPWM method for reducing conducted EMI in single switch three phase discontinuous conduction mode boost converter is presented. The more white noise is injected, the more conducted EMI is reduced. But output-voltage is not sufficiently regulated. This is the reason why carrier frequency selection topology is proposed. In the case of carrier frequency selection, output-voltage of steady state and transient state is fully regulated. A RPWM control method was proposed in order to smooth the switching noise spectrum and reduce it's level. Experimental results are verified by converter operating at 300v/1kW with $5%{\sim}30%$ white noise input. Spectrum analysis is performed on the Phase current and the CM noise voltage. The former is measured with Current Probe and the latter is achieved with LISN, which are connected to the spectrum analyzer respectively.

  • PDF