• 제목/요약/키워드: Differential transcription

검색결과 167건 처리시간 0.025초

Sonicated Protein Fractions of Mycoplasma hyopneumoniae Induce Inflammatory Responses and Differential Gene Expression in a Murine Alveolar Macrophage Cell Line

  • Damte, Dereje;Lee, Seung-Jin;Birhanu, Biruk Tesfaye;Suh, Joo-Won;Park, Seung-Chun
    • Journal of Microbiology and Biotechnology
    • /
    • 제25권12호
    • /
    • pp.2153-2159
    • /
    • 2015
  • Mycoplasma hyopneumoniae is known to cause porcine enzootic pneumonia (EP), an important disease in swine production. The objective of this study was to examine the effects of sonicated protein fractions of M. hyopneumoniae on inflammatory response and gene expression in the murine alveolar macrophage MH-S cell line. The effects of sonicated protein fractions and intact M. hyopneumoniae on the gene expression of cytokines and iNOS were assessed using RT-PCR. The Annealing Control Primer (ACP)-based PCR method was used to screen differentially expressed genes. Increased transcription of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, COX-2, and iNOS mRNA was observed after exposure to the supernatant (SPT), precipitant (PPT), and intact M. hyopneumoniae protein. A time-dependent analysis of the mRNA expression revealed an upregulation after 4 h for IL-6 and iNOS and after 12 h for IL-1β and TNF-α, for both SPT and PPT; the fold change in COX-2 expression was less. A dose- and time-dependent correlation was observed in nitrite (NO) production for both protein fractions; however, there was no significant difference between the effects of the two protein fractions. In a differential gene analysis, PCR revealed differential expression for nine gene bands after 3 h of stimulation — only one gene was downregulated, while the remaining eight were upregulated. The results of this study provide insights that help improve our understanding of the mechanisms underlying the pathogenesis of and macrophage defenses against M. hyopneumoniae assault, and suggest targets for future studies on therapeutic interventions for M. hyopneumoniae infections.

Differential Expression of Taste Receptors in Tongue Papillae of DBA Mouse

  • Choi, Ha-Jung;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
    • /
    • 제41권1호
    • /
    • pp.25-32
    • /
    • 2016
  • The tongue has 4 kinds of papillae, which are filiform, fungiform (FU), foliate (FO) and circumvallate papilla (CV). Tongue papillae except filiform papilla include taste buds. The papillae differ in taste sensitivities, likely due to differential expression of taste receptors. In this study, we evaluated differences in the expression levels of taste receptors in FU, FO and CV. Male DBA2 mice, 42-60 days old, were used in the study. Messenger RNAs were extracted from the murine epithelial tissues including FU, FO and CV. Cloned DNAs were synthesized by reverse transcription. Quantitative PCRs (qPCRs) were performed to determine mRNA expression levels of taste receptors. Results of qPCR revealed that the relative expression levels and patterns were different among FU, FO and CV. All three type 1 taste receptors were expressed FU, FO and CV at varying relative expression levels. All 35 kinds of type 2 taste receptors showed higher expression in FO and CV than in FU. Tas2r108 and Tas2r137 showed the two highest expression levels in all tested papillae. The differential expression levels and patterns of taste receptors among the three papillae could contribute to the different physiological sensitivities by tongue areas. Additional studies such as in situ hybridization or taste receptor cell activity recording is necessary to elucidate the functional relationship between expression levels of taste receptors and taste sensitivity.

Suppression Subtractive Hybridization Identifies Novel Transcripts in Regenerating Hydra littoralis

  • Stout, Thomas;McFarland, Trevor;Appukuttan, Binoy
    • BMB Reports
    • /
    • 제40권2호
    • /
    • pp.286-289
    • /
    • 2007
  • Despite considerable interest in the biologic processes of regeneration and stem cell activation, little is known about the genes involved in these transformative events. In a Hydra littoralis model of regeneration, we employed a rapid shotgun suppression subtractive hybridization strategy to identify genes that are uniquely expressed in regenerating tissue. With an adaptor-PCR based technique, 16 candidate transcripts were identified, 15 were confirmed unique to mRNA isolated from hydra undergoing regeneration. Of these, 6 were undescribed in GenBank and allied expressed sequence tag (EST) databases (GenBank + EMBL + DDBJ + PDB and the Hydra EST database). BLAST analysis of these sequences identified remarkably similar sequences in anonymous ESTs found in a wide variety of animal species.

Differentiation between Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus in Formalin-fixed Paraffin-embedded Tissues by Multiplex RT-nested PCR and Comparison with in situ Hybridization

  • Jung, Kwon-il;Kim, Jung-hyun;Chae, Chan-hee
    • 한국수의병리학회:학술대회논문집
    • /
    • 한국수의병리학회 2003년도 추계학술대회초록집
    • /
    • pp.27-27
    • /
    • 2003
  • Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) infections are considered difficult to distinguish clinically and histopathologically. Prompt differentiation between PEDV- and TGEV-associated enteritis would greatly facilitate the management of disease in countries where PEDV and TGEV are epizootic. Rapid differential diagnosis and treatment are crucial to reducing mortality and morbidity from PEDV- and TGEV-induced enteritis in piglets. The objective for this study was to develop a protocol to differentiate between PEDV and TGEV directly from formalin-fixed, paraffin-embedded tissue, using a multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR) assay. (omitted)

  • PDF

일차 배양 랫드 혈관 평활근 세포에서 Prostaglandin 생성에 미치는 Bacterial Lipopolysaccharide의 작용 특성 (Effects of Bacterial Lipopolysaccharide on Prostaglandin Production in Primary Cultured Rat Vascular Smooth Muscle Cells)

  • 이수환
    • 한국식품위생안전성학회지
    • /
    • 제11권4호
    • /
    • pp.227-234
    • /
    • 1996
  • This study was designed to characterize endotoxin-induced prostaglandin production in primary cultured rat vascular smooth muscle cells (VSMC). The time course for prostaglandin synthesis in lipopolysaccharide (LPS)-stimulated VSMC showed that the maximum production was reached in 12 hours. LPS induced prostaglandin H2 synthase (PGHS) activity in VSMC and the time course profile in the changes of PGHS activity paralleled that of total prostaglandin production. Differential treatment showed that 4 hours' exposure to LPS was enough for the maximum effect on the prostaglandin production and this effect was completely inhibited by the co-treatment of actinomycin D, a transcription inhibitor. These results suggest that LPS effect might be determined within 4 hours. Actinomycin D increased PGHS activity without affecting prostaglandin production if added 4 hours after LPS treatment. On the other hand, cyclogeximide, a translation inhibitor, augmented LPS-induced prostaglandin production if treated during first four hours, but it inhibited LPS-induced PGHS activity regardless of treatment schedule. These results suggest the existence of multiple regulating mechanisms in the LPS-induced prostaglandin synthesis.

  • PDF

한국산 작약에서 분리한 바이로이드 유사 RNA 분자의 확인과 유전자 분석 (Detection and Genomic Analysis of Viroid-like RNA Molecules Isolated from Korean Peonies)

  • 정동수;김무인;이재열
    • 한국식물병리학회지
    • /
    • 제13권2호
    • /
    • pp.113-117
    • /
    • 1997
  • Low moleuclar weight (LMW) RNAs were isolated form Korean peonies which expressed symptoms of stunt and epinasty. The LMW plant RNAs were purified by Qiagen column chromatography which could separate viroid specific nucleic acid at differential salt concentration. After the inoculation of the purified RNAs from the peonies, the inoculated tomatoes (cv. Rutgers) expressed the symptoms of stunt and epinasty. Also the same molecular weight RNAs with viroid-like RNAs were isolated from the inoculated tomatoes. Double-stranded cDNA were synthesized by the methods of reverse transcription (RT) and polymerase chain reaction (PCR) with the purified RNA and primers. The same cDNAs associated with viroid-like RNAs wre cloned from the inoculated tomatoes. The cDNA has been sequenced and its 375-nucleotides were arranged into secondary structure. The cloned cDNA showed 47~54% homology compared with other viroids. The sequence homology of the cloned cDNA were partially high with plant genomic RNAs.

  • PDF

Several Genes Expressed During Morphogenesis of Lentinus edodes(ImHyup-1)

  • Lee, Sang-Sun;Hong, Sung-Woon;Kim, Seung-Hae;Kim, Bong-Cheol
    • Mycobiology
    • /
    • 제29권3호
    • /
    • pp.135-141
    • /
    • 2001
  • Differential display of reverse transcription(DDRT)-PCR was conducted to have a profile of the differentially expressed genes during the formation of fruiting body of Lentinus edodes. The lines of L. edodes(ImHyup-1) employed were cultivated in the artificial blocks of sawdust, and the fruiting body was induced from the mycelia or the mass protruded from the brown surface of the sawdust blocks. RNAs were prepared from the four different developmental stages; mycelial, primordial, and stipes and pileus of fruiting body. The fragments of cDNA were synthesized from the combinations of the arbitrary primers and 3' one anchored Oligo-dT primer. Twelve combinations using the primers have been tested, and among them nineteen bands were identified as differentially expressed. Those genes were further analyzed by DNA sequencing and followed by homology search. Characterization of one clone was conducted as a preliminary data and more are under investigation.

  • PDF

pH Response Pathways in Fungi: Adapting to Host-derived and Environmental Signals

  • Selvig, Kyla;Alspaugh, J. Andrew
    • Mycobiology
    • /
    • 제39권4호
    • /
    • pp.249-256
    • /
    • 2011
  • Microorganisms are significantly affected when the ambient pH of their environment changes. They must therefore be able to sense and respond to these changes in order to survive. Previous investigators have studied various fungal species to define conserved pH-responsive signaling pathways. One of these pathways, known as the Pal/Rim pathway, is activated in response to alkaline pH signals, ultimately targeting the PacC/Rim101 transcription factor. Although the central signaling components are conserved among divergent filamentous and yeast-like fungi, there is some degree of signaling specificity between fungal species. This specificity exists primarily in the downstream transcriptional targets of this pathway, likely allowing differential adaptation to species-specific environmental niches. In this review, the role of the Pal/Rim pathway in fungal pH response is discussed. Also highlighted are functional differences present in this pathway among human fungal pathogens, differences that allow these specialized microorganisms to survive in the various micro-environments of the infected human host.

Comparative proteome analysis of seeds of proso millet (Panicum miliaceum) cultivars

  • Roy, Swapan Kumar;Kwon, Soo Jeong;Park, Hyeong-Jun;Yu, Je-Hyeok;Sarker, Kabita;Cho, Seong-Woo;Jung, Tae-Wook;Park, Cheol-Ho;Woo, Sun-Hee
    • 한국작물학회:학술대회논문집
    • /
    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
    • /
    • pp.120-120
    • /
    • 2017
  • Since the composition of proteins from the Korean cultivars of Proso millet is unknown; thereby, the present study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from the millet seeds of various cultivars, were investigated using proteomic techniques as 2D electrophoresis coupled with mass fingerprinting. The 1152 (differentially expressed) proteins were detected on 2-D gel. Among them, 26 reproducible protein spots were analyzed by MALDI-TOF-TOF mass spectrometry. Out of 26 proteins, 2 proteins were up-regulated towards all cultivars of millet, while 7 proteins were up-regulated and 13 proteins were down-regulated against only one cultivar. However, abundance in most identified protein species, associated with metabolism, transcription and transcription was significantly enhanced, while that of another protein species involved in polysaccharide metabolism, stress response and pathogenesis were severely reduced. Taken together, the results observed from the study suggest that the differential expression of proteins from the four cultivars of millet may be cultivar-specific. Taken together, a proteomic investigation of millet seeds from different cultivars, we sought to better understand the genetic variation of millet cultivars representing the future millet research, and the functional categorization of individual proteins on the basis of their molecular function.

  • PDF