• 제목/요약/키워드: Rep protein

검색결과 33건 처리시간 0.024초

DNA Replication is not Required in Re-establishment of HMRE Silencer Function at the HSP82 Yeast Heat Shock Locus

  • Lee, See-Woo;Gross, David S.
    • Journal of Microbiology
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    • 제34권1호
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    • pp.30-36
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    • 1996
  • We have exmained the re-establishment of HIMRE mediated silencing function on the transcriptional activity of yeast heast shock gene HSP82. To test whether the onset of SIR repression can occur in growing cells in the rpesence of a potent inhibitor of DNA replication, HMRa/HSP82 strains with SIR4- and SIR4S$^{+}$ genetic backgrounds were arrested in S phase by incubation of a culture in 200 mM hydroxyurea for 120 min. It was clear that following a 20 minute heat shock, silencing of the HMRa/HSP82 allele in cells pretreated with hydroxyurea does occur in a SIR4-dependen fashion, even though the kinetics of repression appears to be substantially delayed. We also have tested whether re- establishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.s.

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MCF-7 세포주에서$\gamma$선에 의한 세포신호 전달 관련 유전자의 발현 양상의 분석 (Signal Transduction-related Gene Expression Analysis in MCF-7 followed by $\gamma$-radiation)

  • 박지윤;황창일;박웅양;김진규;채영규
    • 환경생물
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    • 제21권1호
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    • pp.52-55
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    • 2003
  • There is considerable evidence that ionizing radiation (IR) mediates checkpoint control, repair and cell death. In this study, we have used a high density microarray hybridization approach to characterize the transcriptional response of human breast carcinoma MCF-7 cell line to ${\gamma}$-radiation, such as 4 Gy 4 hr, 8 Gy 4 hr, and 8 Gy 12 hr. We found that exposure to ${\gamma}$-ray alters by at least a $log_2$ factor of 1.0 the expression of 115 known genes. Of the 66 genes affected by ${\gamma}$-radiation, 49 are down-regulated. In our results, the cellular response to irradiation includes induction of the c-jun and EGR1 early response genes. The present work has examined potential cytoplasmic signaling cascades that transduce IR-induced signals to the nucleus. 40S ribosomal protein s6 kinase modulates the activities of the mitogen activated protein kinase (MAPK) and c-Jun $NH_2$-terminal kinase (JNK1) cascades in human monocytic leukemia (U937/pREP4) cells. 14-3-3 family members are dimeric phosphoserine -binding proteins that participate in signal transduction and checkpoint control pathways.

pVC, a Small Cryptic Plasmid from the Environmental Isolate of Vibrio cholerae MP-1

  • Zhang, Ruifu;Wang, Yanling;Leung, Pak Chow;Gu, Ji-Dong
    • Journal of Microbiology
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    • 제45권3호
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    • pp.193-198
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    • 2007
  • A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

한국산 길조류에서 추출한 Fucoidan의 황산기에 따른 항암작용 (Increased Anticancer Activity by the Surfated Funcoidan from Korean Brown Seaweeds)

  • 박장수;김안드레;김은희;서홍숙;최원철
    • 대한화학회지
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    • 제46권2호
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    • pp.151-156
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    • 2002
  • Fucoidan은 갈조류에 다량 함유되어있는 다당류로서 항암작용, 항혈응고작용, 항혈전작용, 항염증작용, 항virus작용과 같은 생리활성작용력을 가지고 있어 최근 많은 연구가 수행되어지고 있다. 본 논문은 갈조류에서 fucoidan을 추출 하여 그 항암활성을 SV40 DNA replication assay, RPA-ssDNA binding assay, 그리고 MCF7 cell growth inhibition assay를 이용하여 알아보았으며 이 항암활성은 fucoidan의 황산기가 중요한 역할을 하고 있음을 알아내었다. 화학적으로 황산기의 함유량을 증가시킨 fucoidan이 황산기의 함유량을 제거시킨 fucoidan보다 항암활성이 뛰어났고, 이는 RPA의 ssDNA 결함력을 떨어 EM리기 때문이라 예측되어 진다. 본 연구는 한국산 갈조류에서 추출한 황산기를 함유한 fucoidan의 항암활성능력을 보여주었다.

Recommendation of Nitrogen Topdressing Rates at Panicle Initiation Stage of Rice Using Canopy Reflectance

  • Nguyen, Hung T.;Lee, Kyu-Jong;Lee, Byun-Woo
    • Journal of Crop Science and Biotechnology
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    • 제11권2호
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    • pp.141-150
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    • 2008
  • The response of grain yield(GY) and milled-rice protein content(PC) to crop growth status and nitrogen(N) rates at panicle initiation stage(PIS) is critical information for prescribing topdress N rate at PIS(Npi) for target GY and PC. Three split-split-plot experiments including various N treatments and rice cultivars were conducted in Experimental Farm, Seoul National University, Korea in 2003-2005. Shoot N density(SND, g N in shoot $m^{-2}$) and canopy reflectance were measured before N application at PIS, and GY, PC, and SND were measured at harvest. Data from the first two years(2003-2004) were used for calibrating the predictive models for GY, PC, and SND accumulated from PIS to harvest using SND at PIS and Npi by multiple stepwise regression. After that the calibrated models were used for calculating N requirement at PIS for each of nine plots based on the target PC of 6.8% and the values of SND at PIS that was estimated by canopy reflectance method in the 2005 experiment. The result showed that SND at PIS in combination with Npi were successful to predict GY, PC, and SND from PIS to harvest in the calibration dataset with the coefficients of determination ($R^2$) of 0.87, 0.73, and 0.82 and the relative errors in prediction(REP, %) of 5.5, 4.3, and 21.1%, respectively. In general, the calibrated model equations showed a little lower performance in calculating GY, PC, and SND in the validation dataset(data from 2005) but REP ranging from 3.3% for PC and 13.9% for SND accumulated from PIS to harvest was acceptable. Nitrogen rate prescription treatment(PRT) for the target PC of 6.8% reduced the coefficient of variation in PC from 4.6% in the fixed rate treatment(FRT, 3.6g N $m^{-2}$) to 2.4% in PRT and the average PC of PRT was 6.78%, being very close to the target PC of 6.8%. In addition, PRT increased GY by 42.1 $gm^{-2}$ while Npi increased by 0.63 $gm^{-2}$ compared to the FRT, resulting in high agronomic N-use efficiency of 68.8 kg grain from additional kg N. The high agronomic N-use efficiency might have resulted from the higher response of grain yield to the applied N in the prescribed N rate treatment because N rate was prescribed based on the crop growth and N status of each plot.

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Energy utilization, nutrient digestibility and bone quality of broiler chickens fed Tanzania-type diets in different forms with enzymes

  • Chang'a, Edwin Peter;Abdallh, Medani Eldow;Ahiwe, Emmanuel Uchenna;Al-Qahtani, Mohammed;Mbaga, Said;Iji, Paul Ade
    • Journal of Animal Science and Technology
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    • 제61권4호
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    • pp.192-203
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    • 2019
  • A study was conducted to determine the influence of feed form and microbial enzyme supplementation on energy utilization, bone quality, and amino acid and mineral digestibility of broiler chickens. Four hundred and eighty Ross 308, day-old broiler chickens were randomly assigned to eight diets formulated from commonly used ingredients in Tanzania. A 2 (pellet or mash) ${\times}$ 4 (control, Axtra XB, Quantum Blue (QB) and Axtra XB + QB enzyme) factorial array in a completely randomized design having six replicates per treatment (10 birds per replicate) was used. Birds were raised in climate-controlled rooms in a 3-phase; starter (0-10 days), grower (11-24 days) and finisher (25-35 days). Apparent metabolizable energy (AME), metabolizable energy intake, net energy of production, energy retained as protein (REp), and efficiency of metabolizable energy use for energy and protein retention were higher (p < 0.05) in birds fed pelleted diets. The AME and REp was higher (p < 0.05) with enzyme supplementation. Ash content, weight, length, width and breaking strength of tibia bones were highest (p < 0.05) in birds on pelleted diets. Tibia bone traits were improved (p < 0.05) when enzymes were included, particularly in a combination of QB and Axtra XB. However, potassium, magnesium, and zinc contents were highest (p < 0.05) when QB was supplemented. Digestibility of all amino acids was higher (p < 0.05) in birds supplied with pellets and with enzyme supplementation for most amino acids, except for serine. There was a positive interaction (p < 0.05) between feed form and enzymes on lysine and phenylalanine digestibility. Digestibility of Ca, P, K, S, Zn, and Fe was higher (p < 0.05) in birds fed pelleted diets, while those on mashed diets had higher (p < 0.05) digestibility of Cu and B. The digestibility of P, K, and Zn was highest (p < 0.001) when QB was added, while Ca, P, S, and B digestibility was highest when a combination of Axtra XB + QB was applied. Pelleted diets with or without enzymes improved energy utilization, digestibility of amino acids, and minerals, and increased bone strength in broiler chickens.

Schizosaccharomyces pombe nup97, which Genetically Interacts with mex67, is Essential for Growth and Involved in mRNA Export

  • Cho, Hyun-Jin;Hwang, Duk-Kyung;Jung, Sun-Im;Yoon, Jin-Ho
    • Journal of Microbiology
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    • 제45권4호
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    • pp.344-349
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    • 2007
  • We have isolated previously three synthetic lethal mutants in Schizosaccharomyces pombe, which genetically interact with mex67, in order to identify the genes involved in mRNA export. A novel nup97 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex3. The nup97 gene contains one intron and encodes an 851 amino-acid protein that is similar to nucleoporins, Nppl06p in S. pombe and Nic96p in Saccharomyces cerevisiae. The nup97 gene is essential for vegetative growth, and nup97 null mutant harboring pREP41X-Nup97 showed $poly(A)^+$ RNA export defect when expression of nup97 is repressed in the presence of thiamine. These results suggest that nup97 is involved in mRNA export from the nucleus to cytoplasm.

Identification and Characterization of the Replication Region of Virulence Plasmid pEIB202 in Edwardsiella piscicida

  • Chang, Xinyue;Teng, Chengli;Wu, Haizhen;Ye, Jiang;Wang, Qiyao;Zhang, Huizhan
    • Journal of Microbiology and Biotechnology
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    • 제29권8호
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    • pp.1273-1280
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    • 2019
  • Edwardsiella piscicida is the causative agent of edwardsiellosis, which has caused enormous economic losses worldwide. In our previous research, an attenuated live vaccine known as WED and based on the virulent strain E. piscicida EIB202 can effectively protect turbots against edwardsiellosis via intraperitoneal injection, while vaccination by immersion exhibits a weaker effect. During the development of the immersion vaccine, we surprisingly found the counts of ${\Delta}pEIB202/EIB202$ colonized on zebrafish were 100 times lower than those of EIB202. However, pEIB202 carries 53 predicted ORFs and has several copies in E. piscicida EIB202, impeding the study of its function. Thus, the replication region is located at a 1,980 bp fragment (from 18,837 to 20,816 bp), containing a transcriptional repressor and a replication protein. Moreover, the minimal replication plasmid, named pRep-q77, has low copies in both E. coli and E. piscicida, but is more stable in E. piscicida than in E. coli. This work lays a foundation for further examination of the function of the virulence plasmid pEIB202.

Complete genome sequence of bacteriocin-producing Ligilactobacillus salivarius B4311 isolated from fecal samples of broiler chicken with anti-listeria activity

  • Subin Han;Arxel G. Elnar;Chiwoong Lim;Geun-Bae Kim
    • Journal of Animal Science and Technology
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    • 제66권1호
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    • pp.232-236
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    • 2024
  • Ligilactobacillus is a genus of Gram-positive lactobacilli commonly found in the intestinal tracts of vertebrates. It has been granted a Qualified Presumption of Safety (QPS) status from the European Food Safety Authority (EFSA). One specific strain, Ligilactobacillus salivarius B4311, was isolated from fecal samples of broiler chickens from a farm associated with Chung-Ang University (Anseong, Korea). This strain was observed to have inhibitory effects against Listeria monocytogenes. In this paper, we present the complete genome sequence of Lig. salivarius B4311. The whole genome of strain B4311 comprises 2,071,255 bp assembled into 3 contigs representing a chromosome, repA-type megaplasmid, and small plasmid. The genome contains 1,963 protein-coding sequences, 22 rRNA genes, and 78 tRNA genes, with a guanine + cytosine (GC) content of 33.1%. The megaplasmid of strain B4311 was found to contain the bacteriocin gene cluster for salivaricin P, a two-peptide bacteriocin belonging to class IIb.

Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • 제33권2호
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.