• Title/Summary/Keyword: Promoters

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Effects on the growth promotion of lactic acid bacteria, antimicrobial effect and antioxidant activity of natural materials

  • Woo Jin Ki;Gereltuya Renchinkhand;Tae-Hwan Kim;Myoung Soo Nam
    • Korean Journal of Agricultural Science
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    • v.51 no.3
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    • pp.341-350
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    • 2024
  • According to various negative effects of antibiotic growth promoters, the effects of nature-derived alternatives to antibiotic growth promoters are mainly investigated in livestock industry these days. The effects of nature-derived alternatives to antibiotic growth promoters are mainly investigated in livestock industry these days. This study was conducted to estimate the efficiency of natural-derived alternatives to antimicrobial and antioxidant such as oregano oil (OGO), sulfide type antimicrobial peptides derived from Bacillus or Yeast (SAPBY), antimicrobial peptides derived from yeast (APY). The lactic acid bacteria growth effects were found to be minimal. The antibacterial activity of SAPBY was minimal, but OGO and APY showed antibacterial activity against Escherichia coli and Salmonella Typhimurium, and APY also showed antibacterial activity against Staphylococcus. Antioxidant effects were observed in all SAPBY, OGO, and APY, and when comparing the radical scavenging rate at 10 mg·mL-1 for each sample, OGO showed the highest at 84.9%, followed by SAPBY at 48.7%. Therefore, the substances that can be used as natural antibiotics are SAPBY, APY, and OGO. It is believed that adding these substances to feed and feeding them to livestock will greatly contribute to improving the health of livestock and the productivity of safe livestock products. In further, research focused on various nature-derived alternatives to antibiotic growth promoter in livestock industry is needed.

Evaluation of Novel Constitutive Expression Vectors Equipped with Mined Promoters from Metagenome (메타게놈에서 발굴한 프로모터를 장착한 새로운 항시발현 벡터의 가치평가)

  • Han, Sang-Soo;Kim, Geun-Joong
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.260-267
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    • 2008
  • The choice of expression vector is very important for industrial production of proteins. Therefore, the systematic mining of promoters over a wider range of genetic resource and/or host is required. We previously reported a novel bidirectional reporting system (pBGR) for the isolation of promoters from metagenome and screened useful promoters that functioned constitutively in E. coli under general culture conditions. Among them, three promoter sequences including each upstream region were amplified by PCR and used to construct new expression vectors. To facilitate subcloning, a multi-cloning site was incorporated into the downstream region of the revere primer sequence. At these sites, GFP, esterase and $\beta$-glucosidase were subcloned and analyzed the constitutive expression ability of new promoter in terms of protein solubility and expression level. As a result, these vectors expressed the proteins constitutively to a level of $2{\sim}3%$ of the total cell protein in soluble fraction (>80 %). This study suggested that excavation of metagenomic promoters for construction of expression vector in a certain strain could provide a way for the development of the expression systems.

Construction of High Sensitive Detection System for Endocrine Disruptors with Yeast n-Alkane-assimilating Yarrowia lipolytica

  • Cho, Eun-Min;Lee, Haeng-Seog;Eom, Chi-Yong;Ohta, Akinori
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1563-1570
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    • 2010
  • To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

Linker Scanning Analysis of the BPV-1 Upstream Regulatory Region

  • Kim, Hee-Dai;Rho, Jae-Rang;Choe, Joon-Ho
    • BMB Reports
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    • v.28 no.4
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    • pp.368-373
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    • 1995
  • The upstream regulatory region (URR) of bovine papillomavirus type 1 (BPV) contains promoters and a conditional transcriptional enhancer that is trans-activated by the viral E2 protein. After deleting the 5' and 3' ends of BPV URR, BamHI linkers were inserted into several positions of BPV URR without causing an addition or a deletion of URR sequences. Most linker scanning mutations did not show any effects on the transcription of P7940 and P89 promoters in BPV URR. However, several mutants showed reduced transcriptional activities. Based on our results we found that the AP-2 and Sp1 binding sites were important for basal level transcription of BPV URR in the absence of the E2 protein and that the CTF/NF-1 site is dispensable for E2 transactivation of BPV URR transcription.

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Effect of Cyclic AMP on the Two Promoters of Escherichia coli Thioredoxin Gene

  • Sa, Jae-Hoon;Fuchs, James A.;Lim, Chang-Jin
    • BMB Reports
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    • v.30 no.5
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    • pp.320-325
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    • 1997
  • Thioredoxin is a multi-functional protein which is ubiquitous in microorganisms, animals and plants. Previously, expression of the E. coli thioredoxin gene was found to be negatively regulated by cAMP. In the present study, the effect of cAMP on two separate promoters of the E. coli thioredoxin gene was investigated. Cyclic AMP had a repressible effect on P1 and P1P2 promoter activity of the constructs. This effect was also observed in the cya strain. The P2 promoter construct gave very high -galactosidase activity, and its expression was not affected by exogenous cAMP. It was assumed that a cis-acting negative element, probably the cAMP-CRP binding site, might have been deleted in the P1 promoter construct. Repression of the thioredoxin gene expression by cAMP appeared to be independent of ppGpp.

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Gut Development and Health in the Absence of Antibiotic Growth Promoters

  • Dibner, J.J.;Knight, Chris;Yi, G.F.;Richards, J.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.1007-1014
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    • 2007
  • Acceptance of antibiotic growth promoters (AGP) in agricultural animal production is rapidly disappearing. Both government regulations and consumer preference are driving this change. Producers in any country that seek export markets will be forced to give up AGP if they are to sell to the EU and many other markets. This report will first review the history of AGP use in the animal industry and the concerns about development of antimicrobial resistance. A description of the development and structure of the gut and how it is affected by AGP administration will conclude with results of studies to replace AGP with antimicrobial organic acids.