• 제목/요약/키워드: Strain-life Method

검색결과 364건 처리시간 0.031초

A Novel Negative Regulatory Factor for Nematicidal Cry Protein Gene Expression in Bacillus thuringiensis

  • Yu, Ziquan;Bai, Peisheng;Ye, Weixing;Zhang, Fengjuan;Ruan, Lifang;Yu, Ziniu;Sun, Ming
    • Journal of Microbiology and Biotechnology
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    • 제18권6호
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    • pp.1033-1039
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    • 2008
  • A 3-kb HindIII fragment bearing the cry6Aa2 gene and the adjacent and intergenic regions was cloned from Bacillus thuringiensis strain YBT-1518. Two open reading frames (ORFs), namely, orf1 (termed cry6Aa2) and orf2 that were separated by an inverted-repeat sequence were identified. orf1 encoded a 54-kDa protein that exhibited high toxicity to the plant-parasitic nematode Meloidogyne hapla. The orf2 expression product was not detected by SDS-PAGE, but its mRNA was detected by RT-PCR. The orf2 coexpressed with orf1 at a high level in the absence of the inverted-repeat sequence, whereas, the expression level of otfl was decreased. When orf2 was mutated, the level of orf1 expression was enhanced obviously. In conclusion, the inverted-repeat sequence disturbs orf2 expression, and the orf2 downregulates orf1 expression. This is an example of novel negative regulation in B. thuringiensis and a potential method for enhancing the expression level of cry genes.

청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제 (Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme)

  • 최연희;이준승;배소영;양근재;염규원;조동혁;강옥화;백형석
    • 생명과학회지
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    • 제23권2호
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    • pp.259-266
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    • 2013
  • 혈전용해효소를 생산하는 균주 분리를 위해서 우선 한국, 일본 등지에서 모은 21개의 청국장 시료를 준비하였고 총 268개의 균주를 분리하였다. 이 중에서 1% skim milk가 포함된 nutrient agar 배지에서 protease를 생산하는 bacteria를 분리하였고, 이 결과로 22개의 균주가 분리되었다. 균주들은 apiweb을 통해 생화학적 특성에 근거하여 동정하였다. 또한 세균동정을 위해 16S rRNA 염기서열 분석을 수행하였다. 분리된 대부분의 균주는 Bacillus subtilis와 Bacillus amyloliquefaciens였다. 혈전용해효소의 활성은 fibrin plate 방법에 의해 측정되었고 A2-14, A2-20, C1-05, C1-09, F2-01로 명명된 다섯 균주가 선택되었다. 이중에서 A2-20 균주는 강한 혈전용해 활성을 보였고 동정결과 Bacillus amyloliquefaciens에 가까웠다. A2-20 균주에 의해 생산되는 혈전용해효소는 균 상등액을 이용한 gel filtration과 ion exchange chromatography를 거쳐 부분정제 되었다. 부분 정제된 효소의 최적 pH는 7.0이었고, 최적 온도는 $35^{\circ}C$였다. 정제된 단백질의 분석은 SDS-PAGE와 zymography로 이루어졌다. 이와 더불어 혈전용해효소의 유전자적 분석도 수행되었으며 A2-20 균주가 생산하는 혈전용해효소의 부분적인 염기서열과 유전적 상동성을 보이는 서열을 규명하였다.

Enhanced Production of Galactooligosaccharides Enriched Skim Milk and Applied to Potentially Synbiotic Fermented Milk with Lactobacillus rhamnosus 4B15

  • Oh, Nam Su;Kim, Kyeongmu;Oh, Sangnam;Kim, Younghoon
    • 한국축산식품학회지
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    • 제39권5호
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    • pp.725-741
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    • 2019
  • In the current study, we first investigated a method for directly transforming lactose into galacto-oligosaccharides (GOS) for manufacturing low-lactose and GOS-enriched skim milk (GSM) and then evaluated its prebiotic potential by inoculating five strains of Bifidobacterium spp. In addition, fermented GSM (FGSM) was prepared using a potentially probiotic Lactobacillus strain and its fermentation characteristics and antioxidant capacities were determined. We found that GOS in GSM were metabolized by all five Bifidobacterium strains after incubation and promoted their growth. The levels of antioxidant activities including radical scavenging activities and 3-hydroxy-3-methylglutaryl-CoA reductase inhibition rate in GSM were significantly increased by fermentation with the probiotic Lactobacillus strain. Moreover, thirty-nine featured peptides in FGSM was detected. In particular, six peptides derived from ${\beta}$-casein, two peptides originated from ${\alpha}s_1$-casein and ${\kappa}$-casein were newly identified, respectively. Our findings indicate that GSM can potentially be used as a prebiotic substrate and FGSM can potentially prevent oxidative stress during the production of synbiotic fermented milk in the food industry.

Damage detection of 3D printed mold using the surface response to excitation method

  • Tashakori, Shervin;Farhangdoust, Saman;Baghalian, Amin;McDaniel, Dwayne;Tansel, Ibrahim N.;Mehrabi, Armin
    • Structural Engineering and Mechanics
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    • 제75권3호
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    • pp.369-376
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    • 2020
  • The life of conventional steel plastic injection molds is long but manufacturing cost and time are prohibitive for using these molds for producing prototypes of products in limited numbers. Commonly used 3D printers and rapid prototyping methods are capable of directly converting the digital models of three-dimensional solid objects into solid physical parts. Depending on the 3D printer, the final product can be made from different material, such as polymer or metal. Rapid prototyping of parts with the polymeric material is typically cheaper, faster and convenient. However, the life of a polymer mold can be less than a hundred parts. Failure of a polymeric mold during the injection molding process can result in serious safety issues considering very large forces and temperatures are involved. In this study, the feasibility of the inspection of 3D printed molds with the surface response to excitation (SuRE) method was investigated. The SuRE method was originally developed for structural health monitoring and load monitoring in thin-walled plate-like structures. In this study, first, the SuRE method was used to evaluate if the variation of the strain could be monitored when loads were applied to the center of the 3D printed molds. After the successful results were obtained, the SuRE method was used to monitor the artifact (artificial damage) created at the 3D printed mold. The results showed that the SuRE method is a cost effective and robust approach for monitoring the condition of the 3D printed molds.

Trehalose에 의한 Rahnella aquatilis AY2000 균주가 생산하는 항효모성 물질의 활성보호 (Trehalose Protects Activity of Anti-Yeast Substance Produced by Rahnella aquatilis Strain AY2000)

  • 강민정;이종환;이복규;김광현
    • 한국미생물·생명공학회지
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    • 제37권4호
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    • pp.361-364
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    • 2009
  • AYS(Anti-yeast substance)는 열에 의해 항효모 활성이 쉽게 약화되는 물질로서 trehalose의 사용이 AYS의 열에 대한 불안정성을 어느 정도 극복할 수 있을 것인지를 조사하기 위해 trehalose가 첨가된 AYS와 trehalose가 첨가되지 않은 AYS에 대한 항효모 활성을 비교 측정하였다. 그 결과 trehalose가 첨가된 AYS는 고온($50-70^{\circ}C$)에서 단시간(2 hr) 동안 열처리한 경우에 trehalose가 함유되지 않은 AYS에 비해 그 활성이 비교적 잘 유지되었다. 따라서 trehalose를 AYS에 첨가하면 열처리 후에도 AYS의 항효모 활성을 다소 보호할 수 있다.

인장법에 의한 박판 판계 용접부의 잔류 응력 거동 특성에 관한 연구 (A Study on the Behavior Characteristics of Residual Stress of the Thin Butt Weldment by Mechanical Tensioning Method)

  • 김하근;김경규;신상범
    • 대한용접접합학회:학술대회논문집
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    • 대한용접접합학회 2010년도 춘계학술발표대회 초록집
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    • pp.57-57
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    • 2010
  • For thin panel welded structure, the various welding distortions were found due to the low resistance against welding deformation. Especially, buckling distortion induced in the thin panel welded structure produce severe problems related to cost in production stage and safety in service life. So, many researches including mechanical and thermal tensioning method for preventing the occurrence of buckling distortion in the production stage have been performed. The purpose of this study is to identify the behavior of longitudinal residual stress at the SA butt weldment with thin plate of 6mm thickness under tension load by 3 dimensional FEA. For it, mesh design for 3D FEA was constructed with 20 nodes brick element for butt weldment and 8 nodes shell element for base metal. According to FEA results, the longitudinal compressive strain inducing tensile residual stress at the butt weldment decreased. It was because the compressive thermal strain in way of weldment was reduced by tension load. The control effect of residual stress increased with an increase in tension load. So, if the amount of tension load applied to the weldment exceeds 1.5 times of longitudinal shrinkage force, the amount of longitudinal residual stress decreased below the critical value inducing the buckling distortion at the SA butt weldment. Its validity was verified by experiment.

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Identification and Distribution of Bacillus Species in Doenjang by Whole-Cell Protein Patterns and 16S rRNA Gene Sequence Analysis

  • Kim, Tae-Woon;Kim, Young-Hoon;Kim, Sung-Eon;Lee, Jun-Hwa;Park, Cheon-Seok;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • 제20권8호
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    • pp.1210-1214
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    • 2010
  • Many bacteria are involved in the fermentation of doenjang, and Bacillus species are known to perform significant roles. Although SDS-PAGE has been frequently used to classify and identify bacteria in various samples, the microbial diversity in doenjang has not yet been investigated. This study aims to determine the identity and distribution of dominant Bacillus species in doenjang using SDS-PAGE profiles of whole-cell proteins and 16S rRNA gene sequencing. Reference Bacillus strains yielded differential SDS-PAGE banding patterns that could be considered to be highly specific fingerprints. Grouping of bacterial strains isolated from doenjang samples by whole-cell protein patterns was confirmed by analysis of their 16S rRNA gene sequences. B. subtilis was found to be the most dominant strain in most of the samples, whereas B. licheniformis and B. amyloliquefaciens were less frequently found but were also detected in several samples. The results obtained in this study show that a combined identification method using SDS-PAGE profiles of whole-cell proteins and subsequent 16S rRNA gene sequence analysis could successfully identify Bacillus species isolated from doenjang.

Characterization of the Genes Involved in Induced Systemic Resistance in Cucumber Plants

  • Kim, Mi-Seong;Cho, Song-Mi;Im, Yang-Ju;Kim, Young-Cheol;Yang, Kwang-Yeol;Lee, Myung-Chul;Kim, Kwang-Sang;Cho, Baik-Ho
    • 한국자원식물학회지
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    • 제20권2호
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    • pp.216-219
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    • 2007
  • Root colonization by a rhizobacterium, Pseudomonas chlororaphis O6, elicited induced systemic resistance (ISR) in the leaves of cucumber plants against fungal and bacterial pathogens. To understand the role of unique genes during strain O6-mediated ISR, a suppressive subtractive hybridization method was undertaken and led to isolation of twenty-five distinct genes. The transcriptional levels of all the genes showed an increase much earlier under O6 treatment than in water control plants only after challenge with pathogen, while no difference detected on the plants without pathogen challenge. This suggests that O6-mediated ISR is associated with the priming phenomenon, an enhanced capacity for the rapid and effective activation of cellular defense responses after challenge inoculation.

Serratia marcescens의 Polyphosphate Kinase 유전자 특성 (Characterization of Polyphosphate Kinase Gene in Serratia marcescens)

  • Yang Lark Choi;Seung Jin Lee;Ok Ryul Song;Soo Yeol Chung;Young Choon Lee
    • 생명과학회지
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    • 제10권4호
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    • pp.397-402
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    • 2000
  • 본 연구는 인산 축적능이 뛰어난 균주를 분자 육종하여 생물학적 폐수처리 및 토양의 인산 집적을 해결시키는 산업적 유용한 재료로 이용하기 위한 기초연구를 목표로 하고 있다. Polyphosphate kinase의 ATP의 phosphate를 단리하여 한분자씩 결합시키는 형태로 polyphosphate의 합성반응을 촉매한다. 인산 축적에 관한 대사과정의 분자적 이해를 위하여 Serratia marcescens균주로부터 Southern hybridization방법으로 ppk를 암호하는 유전자를 찾아내어 새조합시킨 pDH3를 구축하였다. pDH3으로부터 ppk를 암호하는 유전자 영역의 4.0 kb 단편을 가진 subclone을 작성하였다.Serratia marcescens의 polyphosphate kinase의 활성은 catabolite repression에 의한 조절을 받았다. 발현멕타에 삽입시킨 재조합 플라스미드를 대장균에 도입시킨 결과, polyphosphate kinase의 효소활성이 크게 증가됨을 확인 하였다. 또한 대량 발현시킨 결과를 SDS-PAGE를 통하여 75 KDa의 발현산물을 확인할 수 있었다.

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분자학적 진단방법을 이용한 bifenazate 저항성 점박이응애 모니터링 (Monitoring of Bifenazate Resistant Two-spotted Spider Mite, Tetranychus urticae Using Molecular Detection Method)

  • 이규리;신윤호;조선란;구현나;최장전;안기수;김길하
    • 농약과학회지
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    • 제15권1호
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    • pp.61-67
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    • 2011
  • 2010년에 청주, 충주-1, 충주-2, 강진, 예산, 그리고 영주에서 채집한 점박이응애의 bifenazate 약제 저항성을 확인 해 본 결과, 강진과 예산 개체군에서 각각 964.5배, 1130배의 높은 저항성을 나타내었다. 그리고 청주, 충주-1, 충주-2, 그리고 영주개제군에서는 낮은 저항성비를 나타내었다. Bifenazate 약제 저항성 점박이응애의 cytb 점 돌연변이인 G126S를 확인해 본 결과, 생물검정에서 높은 저항성을 보인 강진과 예산 개체군에서 G126S 점 돌연변이를 확인 할 수 있었다. 이와 같이, G126S 점 돌연변이는 bifenazate 약제 저항성을 가지는 점박이응애 선별에 아주 유용한 분자진단 마커로 이용될 수 있다. 두 가지 분자진단 방법인 quantitative sequencing(QS)와 PCR amplification of specific alleles(PASA)는 G126S 점 돌연변이를 잘 탐지하였다. 따라서 이러한 방법들은 야외 계통의 bifenazate 약제 저항성 형질 모니터링과 저항성 관리에 효율적으로 이용될 수 있을 것이다.