• Title/Summary/Keyword: Promoters

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Development and Verification of an Optimum Composition Model for a Synbiotic Fermented Milk Using Sequential Quadratic Programming Techniques

  • Chen, Ming-Ju;Chen, Kun-Nan;Lin, Chin-Wen
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1490-1495
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    • 2006
  • The purpose of this research was to develop an optimum composition model for a new synbiotic fermented dairy product with high probiotic cell counts, and to experimentally verify this model. The optimum composition model indicated the growth promoter ratio that could provide the highest growth rate for probiotics in this fermented product. Different levels of growth promoters were first blended with milk to improve the growth rates of probiotics, and the optimum composition model was determined. The probiotic viabilities and chemical properties were analyzed for the samples made using the optimal formula. The optimal combination of the growth promoters for the synbiotic fermented milk product was 1.12% peptides, 3% fructooligosaccharides (FOS), and 1.87% isomaltooligosaccharides (IMO). A product manufactured according to the formula of the optimum model was analyzed, showing that the model was effective in improving the viability of both Lactobacillus spp. and Bifidobacterium spp.

Analysis of Heat Shock Promoters in Hansenula polymorpha: The TPS1 Promoter, a Novel Element for Heterologous Gene Expression

  • Amuel, Carsten;Gellissen, Gerd;Cor;Suckow, Manfred
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.247-252
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    • 2000
  • The strength and regulatory characteristics of the heat-inducible HSA1, HSA2 and TPS1 promoters were compared with those of the well-established, carbon source-regulated FMD promoter in a Hansenula polymorpha-based host system in vivo. In addition, the Saccharomyces cerevisiae-derived ADH1 promoter was analysed. While ADH1 promoter showed to be of poor activity in the foreign host, the strength of the heat shock TPS1 promoter was found to exceed that of the FMD promoter, which at present is considered to be the strongest promoter for driving heterologous gene expression in H. polymorpha.

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Machine Learning Applied to Uncovering Gene Regulation

  • Craven, Mark
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2000.11a
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    • pp.61-68
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    • 2000
  • Now that the complete genomes of numerous organisms have been ascertained, key problems in molecular biology include determining the functions of the genes in each organism, the relationships that exist among these genes, and the regulatory mechanisms that control their operation. These problems can be partially addressed by using machine learning methods to induce predictive models from available data. My group is applying and developing machine learning methods for several tasks that involve characterizing gene regulation. In one project, for example, we are using machine learning methods to identify transcriptional control elements such as promoters, terminators and operons. In another project, we are using learning methods to identify and characterize sets of genes that are affected by tumor promoters in mammals. Our approach to these tasks involves learning multiple models for inter-related tasks, and applying learning algorithms to rich and diverse data sources including sequence data, microarray data, and text from the scientific literature.

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Multiple shRNA expressing vector enhances efficiency of gene silencing

  • Song, Jun;Giang, An;Lu, Yingchun;Pang, Shen;Chiu, Robert
    • BMB Reports
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    • v.41 no.5
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    • pp.358-362
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    • 2008
  • RNA interference (RNAi) is the process of sequence-specific gene silencing. However, RNAi efficiency still needs to be improved for effective inhibition of target genes. We have developed an effective strategy to express multiple shRNAs (small hairpin RNA) simultaneously using multiple RNA Polymerase III (Pol III) promoters in a single vector. Our data demonstrate that multiple shRNAs expressed from Pol III promoters have a synergistic effect in repressing the target gene. Silencing of endogenous cyclophilin A (CypA) or key HIV viral genes by multiple shRNAs results in significant inhibition of the target gene.

Secretion Capacity Limitations of the Sec Pathway in Escherichia coli

  • Mergulhao, Filipe J.M.;Monteiro, Gabriel A.
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.128-133
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    • 2004
  • The secretion capacity of two E. coli strains (JM109 and AF1000) was evaluated through the expression of two human proinsulin fusion proteins using the translocation signal sequence from Staphylococcal protein A (SpA). Although a 7 to 11-fold difference in the expression levels was attained by the use of different promoters (SpA and malK promoters) and copy-number vectors (700 and 50 copies per cell), the maximum translocation rates for all the systems were around 140,000 amino acids $cell^{-1} min^{-1}$. Moreover, the secretion capacity was found to be independent of the size of the exiting peptide and its translational rate.

Cloning of Promoters from Alkali-tolerant Bacillus sp. (알카리 내성 Bacillus속 Promoter의 Cloning)

  • 유주현;구본탁;공인수;정용준;박영서
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.126-130
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    • 1988
  • Promoters of an alkali-tolerant Bacillus sp. isolated from soil have been cloned in Bacillus subtilis using promoter probe vector pPL703. The CAT specific activity of a clone harboring the strongest promoter activity among these transformants was 8.01. This activity was 2.5 times higher than that of Bacillus subtilis harboring expression vector pPL708 and was increased after the end of the logarithmic growth phase. In the 2.8kb of inserted DNA fragment, BamHI and Sal I recognition sites were located.

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High-Resolution Microarrays for Mapping Promoter Binding sites and Copy Number Variation in the Human Genome

  • Albert Thomas
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.125-126
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    • 2006
  • NimbleGen has developed strategies to use its high-density oligonucleotide microarray platform (385,000 probes per array) to map both promoter binding sites and copy number variation at very high-resolution in the human genome. Here we describe a genome-wide map of active promoters determined by experimentally locating the sites of transcription imitation complex binding throughout the human genome using microarrays combined with chromatin immunoprecipitation. This map defines 10,567 active promoters corresponding to 6,763 known genes and at least 1,196 un-annotated transcriptional units. Microarray-based comparative genomic hybridisation (CGH) is animportant research tool for investigating chromosomal aberrations frequently associated with complex diseases such as cancer, neuropsychiatric disorders, and congenital developmental disorders. NimbleGen array CGH is an ultra-high resolution (0.5-50 Kb) oligo array platform that can be used to detect amplifications and deletions and map the associated breakpoints on the whole-genome level or with custom fine-tiling arrays. For whole-genome array CGH, probes are tiled through genic and intergenic regions with a median probe spacing of 6 Kb, which provides a comprehensive, unbiased analysis of the genome.

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Impact of SV40 T antigen on two multiple fission microalgae species Scenedesmus quadricauda and Chlorella vulgaris

  • Gomaa, Ahmed E.;Yang, Seung Hwan
    • International journal of advanced smart convergence
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    • v.7 no.1
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    • pp.48-63
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    • 2018
  • The combination of Simian Virus40 (SV40)'s large T antigen with its replication origin is commonly used in molecular studies to enhance the expression of heterogeneous genes through multiplying the plasmid copy number. There are no reports related to the impact of the SV40 T antigen on plant, multiple fissional, cell-type. This study explores the response of two multiple-fission microalgal cells, Scenedesmus quadricauda and Chlorella vulgaris, to the expression of the T-antigen, with aim of applying SV40 T-antigen to increase the expression efficiency of foreign genes in the two species. Different levels of low-expression have been constructed to control the expression of SV40 T antigen using three heterogenous promoters (NOS, CaMV35S, and CMV). Chlorella cultures showed slowdown in the growth rate for samples harboring the T antigen under the control of CaMV35S and CMV promoters, unlike Scenedesmus cultures which showed no significant difference between samples and could have silenced the expression.

Construction of an expression vector with SV40 DNA in a mammalian cell (SV 40 DNA를 이용한 포유동물의 유전자 운반체 개발)

  • 정민혜;김상해;전희숙;노현모
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.165-172
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    • 1987
  • An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1, 118bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVML-TKp. To test the expression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kinase gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK. These plasmids, pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.

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Heat Inducible Expression of the CDC70 Gene Under the Control of Heat Shock Element in Saccharomyces Cerevisiae

  • Lee, Seok-Jae;Jahng, Kwang-Yeop;Lee, Young-Hoon;Chae, Keon-Sang
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.196-200
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    • 1995
  • In order to express the CDC70 gene of Saccharomyces cerevisiae by heat shock, we have designed heat inducibe hybrid promoters using the Drosophila melanogaster heat shock elements (HSEs). A 220 bp-long upstream fragment of the D. melanogaster hsp70 gene comprised of four HSEs was placed upstream of the putative proximal TATA box of the CDC70 gene. Hybrid promoters containing different fusion joints were tested for their ability to drive the CDC70 gene expression by heat shock. The results showed that the HSEs of D. melanogaster conferred the heat-induced CDC70 gene expression, but the heat inducibility was much lower than that in D. melanogaster.

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