• 식품기술
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• 제17권4호
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• pp.98-106
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• 2004

Two mutant enzymes of $\gamma$-glutamylcysteine synthetase ($\gamma$-GCS) which catalyzed the synthesis of $\gamma$-glutamylcysteine from L-glutamic acid and L-cysteine in the presence of ATP, were prepared bypoint mutation of $\gamma$-GCS gene with site-directed mutagensis in E. coli. Conformational structuresand catalytic reaction kinetics of mutant enzymes were compared with wild type $\gamma$-GCS afterpurification. The S495F mutant enzyme (serine at 495 residue was substituted with phenylalanine),which had no catalytic activity for $\gamma$-glutamylcysteine synthesis, rarely folded even in neutral pH.However, the mutant A494V (alanine of 494 residue was replaced by valnine) which showed 50 %increase of activity, had a high folding structure. The folding structure of A494V also more stable athigh temperature and extreme pH compared to wild type and S495F. Reaction kinetics of wild typeand A494V were also investigated, Km value of A494V was smaller than that of wild type, while itshowed a little difference at Vmax values. This result evolved that alanine at 494 may be involved inbinding site of substrate rather than catalytic site. In addition, change of catalytic activity by onepoint mutation was highly correlated with the folding structure of enzyme.

• 한국환경농학회지
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• 제38권3호
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• pp.197-204
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• 2019

BACKGROUND: Living modified microorganisms (LMMs) have been focused in two very different aspects of positive and negative effects on ecology and human health. As a model experiment, wild type and a foreign origin gene-harboring modified E. coli strains were subjected to comparison of their metabolomes and potential effects on soil microbiota in the laboratory sets. This study assumes the unintentional release of LMMs and tries to suggest potential effects on the soil microbiota even at minimal settings. METHODS AND RESULTS: Metabolomes from the wild type and LM E. coli were analyzed by NMR and the profiles were compared. In the laboratory soil experiments, the two types of E. coli were added to the soils and monitored for the bacterial community compositions. Those metabolomic profiles did not show significant differences. The microbial community structures from the time series soil DNAs for both the sets using wild type and LMO also did not indicate significant changes, but minor by the addition of foreign organisms regardless of wild or LMO. CONCLUSION: Even if the recombinant microorganism (LMO) is released into the soil environment, the survival of microorganisms in the environment would be one of the major factors for the transfers of foreign genes to other organisms and diffusion into the soil environment.

• 방사선산업학회지
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• 제3권1호
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• pp.39-45
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• 2009

Three mineral phosphate solubilizing (MPS) bacteria where isolated from rhizosphere soil samples of common bean and weed plants. 16S rDNA analysis indicated that the isolate P2 and P3 are closely related to Pantoea dispersa while isolate P4 is closely related to Pantoea terrae. Isolates P2 and P3 recorded $381.60{\mu}g\;ml^{-1}$ and $356.27{\mu}g\;ml^{-1}$ of tricalcium phosphate (TCP) solubilization respectively on 3 days incubation. Isolate P4 recorded the TCP solubilization of $215.85{\mu}g\;ml^{-1}$ and the pH was dropped to 4.44 on 24 h incubation. Further incubation of P4 sharply decreased the available phosphorous to $28.94{\mu}g\;ml^{-1}$ and pH level was raised to 6.32. Gamma radiation induced mutagenesis was carried out at $LD_{99}$ dose of the wild type strains. The total of 14 mutant clones with enhanced MPS activity and 4 clones with decreased activity were selected based on solubilization index (SI) and phosphate solubilization assay. Mutant P2-M1 recorded the highest P-solubilizing potential among any other wild or mutant clones by releasing $504.21{\mu}g\;ml^{-1}$ of phosphorous i.e. 35% higher than its wild type by the end of day 5. A comparative evaluation of TCP solubilization by wild type isolates of Pantoea and their mutants, led to select three MPS mutant clones such as P2-M1, P3-M2 and P3-M4 with a potential to release >$471.67{\mu}g\;ml^{-1}$ of phosphorous from TCP. These over expressing mutant clones are considered as suitable candidates for biofertilization.

• 한국지역사회생활과학회지
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• 제20권2호
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• pp.205-221
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• 2009

My concern was to explore how the Korean 'wild geese families' functioned to maintain 'familyhood' in spite of spatial separation by using a qualitative approach. I used personal narratives from eleven 'geese mothers' living in the United States. Family paths across time were analyzed to understand their complicated nature. Respondents adopted some coping strategies to obtain the flexibility and the stability for relocating their transnational lives. These were 1) communications, 2) relocation of household work, and 3) reinterpretation of 'sacrifice.' It seemed that their family paths become either the continuous type (prolong their stay) or the discontinuous type (not prolong their stay). These were shaped by complex individual, familial, and social forces which affected differently according to the steps of family life cycles. Therefore, this study showed that the Korean 'wild geese family' did not move toward the uni-direction with the same experiences and nature. More importantly, it is noteworthy to acknowledge that the prevalence of 'wild geese family' reveals the dynamic interactive nature of the family, i.e., actively responding to the changes and challenges from the diverse circumstances. It is inferred that the social and cultural factors such as the class mobility, the education system, and the values may influence the family life style.

• 한방비만학회지
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• 제21권1호
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• pp.42-48
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• 2021

Objectives: The purpose of this study was to review the studies of the wild ginseng complex pharmacopuncture related to obesity treatment. Methods: We searched the papers with key words of 'wild ginseng complex pharmacopuncture', 'wild ginseng pharmacopuncture', 'obesity', 'fat', 'weight' in Oriental medicine Advanced Searching Integrated System, KRpia, Koreanstudies Information Service System, Research Information Sharing Service, KoreaMed and PubMed, Scopus. We classified the papers by year, content and study type. Results: There were 7 studies about the wild ginseng pharmacopuncture related to obesity treatment. Cell studies were excluded because they were not related to obesity. Analysis of 3 animal experiments and 4 clinical studies were conducted to describe each research subject, method, and research results. Conclusion: More interest and further research will be needed on wild ginseng pharmacopuncture related to obesity treatment in the Korean medicine to achieve clinical application and to develop treatment protocols for the obesity disease.

• 대한약리학회지
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• 제25권1호
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• pp.115-134
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• 1989

Ganciclovir(GCV)와 Vidarabine(ara-A)을 단독으로 또는 동시에 HSV-1에 작용시켰을때 HSV-1의 복제, DNA합성 및 단백질 합성에 미치는 영향을 관찰할 목적으로 본 연구를 시행하였다. 본 실험에서는 4가지의 다른 HSV-1(Wild type KOS, $VCV^r$, $IUdR^r$, 및 $PAA^r5$)를 사용하였다. Virus복제에 미치는 항 virus약의 효과는 Vero 세포단층 배양에서 Yield reduction assay에 의해서 관찰하였다. 항 virus약의 virus DNA 합성에 미치는 영향은 NaI 밀도 구배초원심침전후 $H^3-$표지 virus DNA의 방사능에 의해서 관찰하였다. Virus 단백질의 합성에 미치는 항 virus약의 효과는 $^{35}S$를 표지한 후 polyacrylamide 겔 전기영동법, 자가방사기록법 그러고 virus bands의 음영농도주사법을 이용, 관찰하여 다음과 같은 결과를 얻었다. 1. GCV는 wild trype HSV-1 KOS와 $PAA^r5$의 복제를 강력하게 억제하였으나 $ACV^r$와 $IUdR^r$는 GCV에 대해서 중등도의 저항을 보였다. ara-A는 실험대상의 모든 HSV-(KOS, $ACV^r$, $IUdR^r$ 및 $PAA^r5$)의 복제에 대해서 거의 비슷하게 억제효과를 보였다. GCV와 ara-A 동시첨가는 KOS와 $PAA^r5$의 복제에 대해서 상승적인 억제효과를 보였고 $ACV^r$와 $IUdR^r5$의 복제에 대해서는 상가작용이하의 억제 효과를 보였다. 2. GCV또는 ara-A는 HSV-1감염 Vero세포에서 virus DNA의 합성을 유의하게 억제하였다. GCV와 ara-A의 동시첨가는 KOS 또는 $PAA^r5$ 감염세포에서 virus DNA 합성을 GCV 또는 ara-A를 단독 첨가하였을때 보다 현저하게 억제하였다. $ACV^r$ 또는 $IUdR^r$ 감염세포에서는 이런 현상을 관찰할 수 없었다. 3. Wild type HSV-1 감염세포에서 virus-단백질의 합성은 GCV, ara-A의 단독 첨가 또는 GCV와 ara-A의 동시첨가에 의해서 변경되지 않았다. Wild type HSV-1 감염말기에 GCV 또는 ara-A 단독 혹은 동시첨가에 의해서 virus 단백질의 합성은 경미하나마 유의성있게 증가하였다. Wild type HSV-1과 $PAA^r5$에 의한 단백질의 합성은 GCV 또는 ara-A 단독 첨가에 의해서 유의성있게 억제되었다. GCV또는 ara-A의 동시첨가는 GCV또는 ara-A를 단독으로 첨가했을 경우보다 단백질의 합성을 더욱 억제하였다. 이상의 실험결과로 보아 GCV와 ara-A의 동시사용은 HSV-1 혹은 ACV저항 DNA polymerase변이주인 $PAA^r5$에 대해서 상승적인 억제작용을 나타냈으며 이 효과는 virus DNA 합성 억제에 의한 것으로 생각된다. ACV저항 thymidine kinase 변이주인 $ACV^r$ 및 $IUdR^r$에 대해서는 ara-A가 유효하였다. 항 virus 약물에 의한 virus 단백질합성의 변화는 virus DNA 합성에 대한 억제효과에 인한 것으로 사려된다.

• BMB Reports
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• 제45권6호
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• pp.342-347
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• 2012

Plant mitochondria possess alternative respiratory pathways mediated by the type II NAD(P)H dehydrogenases and alternative oxidases. Here, E3 SUMO ligase was shown to regulate alternative respiratory pathways and to participate in the maintenance of carbon and nitrogen balance in Arabidopsis. The transcript abundance of the type II NAD(P)H dehydrogenases NDA2 and NDB2 and alternative oxidases AOX1a and AOX1d genes was low in siz1-2 mutants compared to that in wild-type. The addition of nitrate or ammonium resulted in a decrease or an increase in the expression of the same gene families, respectively, in both wild-type and siz1-2 mutants. The amount of free sugar (glucose, fructose and sucrose) was lower in siz1-2 mutants than that in wild-type. These results indicate that low nitrate reductase activity due to the AtSIZ1 mutation is correlated with an overall decrease in alternative respiration and with a low carbohydrate content to maintain the carbon to nitrogen ratio in siz1-2 mutants.

• 한국수정란이식학회지
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• 제29권2호
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• pp.195-200
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• 2014

Endothelin 2 (EDN2) induces follicular rupture by constricting periovulatory follicles. In this study, it was investigated the mechanisms of EDN2 action on follicular rupture with respect of receptor using the conditionally granulosa cell specific EDN2 receptor type A (ETa) KO mice (gcETaKO; $ETa^{flox/-}{\cdot}Amhr2^{Cre}$). It was generated the gcETaKO mice by breeding with $ETa^{flox/-}$ mice after mono-alleic ETa knockout by $ZP3^{Cre}$ and $Amhr2^{Cre}$ mice. Fertility, ovulation and maturation rates of ovulated oocytes after super ovulation were investigated in the gcETaKO mice compared with wild-type mice ($ETa^{flox/flox}$ and $ETa^{flox/-}$) as a control group. In the gcETaKO mice, normal fertility after breeding with male mice was shown compared with wild-type mice. And, there was no significant differences in ovulation rates after super ovulation, however its maturation rates was lower than that of wild type mice. These findings show that EDN2 in follicular rupture for ovulation is related with an other ETa not in granulosa cells. Further studies are needed to investigate how EDN2 is acted in ovarian follicular rupture for ovulation.

• 대한수의학회지
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• 제58권2호
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• pp.95-98
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• 2018

Brucellosis is one of the common zoonoses caused by Brucella abortus (B. abortus). However, little has been reported on factors affecting invasion of B. abortus into host cells. To investigate cell-type dependent invasion of B. abortus, phagocytic RAW 264.7 and THP-1 cells and non-phagocytic HeLa cells were infected with wild-type and mutant B. abortus, and their invasion efficiencies were compared. The invasion efficiencies of the strains were cell-type dependent. Wild-type B. abortus invasion efficiency was greater in phagocytic cells than in epithelial cells. The results also indicated that there are different factors involved in the invasion of B. abortus into phagocytic cells.

• 한국식물병리학회지
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• 제13권3호
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• pp.179-183
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• 1997

십자화과 식물에 검은썩음병을 일으키는 Xanthomonas campestris pv. campestris에 대하여 담배 18품종 중 가장 과민반응을 강하게 나타내는 품종(Nicotiana tabacum cv. TC500)을 과민반응 비기주식물로 선발하였다. NTG 돌연변이를 실시하여 얻은 약 4,500개의 mutant들을 $10^{8}$ cells/ml의 농도로 주사 접종하여 비기주식물인 담배 잎에서 과민반응을 유도하지 않는 HR-mutans(XHN 514-774, XHN 620-831)를 선발하였고, 기주식물인 양배추 잎에서 병원성을 나타내지 않는 비병원성변이주(XPN 1001)를 선발하였다. HR mutant를 기주식물인 양배추에 접종한 결과 병원성을 나타내었고, nonpathogenic mutant를 비기주식물인 담배잎에 접종한 결과 과민반응을 유도하였다. 담배 잎에서 HR-mutant와 wild type의 population를 조사한 결과 wild type은 24시간 이후 급격히 감소하는 반면, HR mutant는 일정수준을 유지하였으나 72시간 이후에 감소하였다. 그리고 mutant들의 protease, cellulase, amylase, polygalacturonate lyase 분비여부를 조사한 결과 wild type과 비슷하게 분비하였다.