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재조합 단백질 과발현을 위한 Bacillus snbtilis 포자형성 변이주의 개발 및 특성 분석
오민규,박승환김병기 한국생물공학회 1994 KSBB Journal Vol.9 No.1
Bacillus subtilis를 재조합 이종 단백질 생산 균주로 만들기 위하여 포자형성 변이주를 만들었다. 균주는 두 개의 프로테아제가 제거된 균주인 DB104로부터 spoOJ와 spoIIG 변이주를 유전자 조작법에 의해 만들고 두 개의 유전자가 모두 제거된 균주도 만들었다. 이에 목적 aprE 유전자를 삽입 벡터 형태로 만들어 integration시킨 뒤 변이주 각각의 형태적인 변화를 투과성 전자현미경으로 살펴 보았다. 각각 변이주의 모습은 이전에 보고된 것과 거의 일치하였으며 spoOJ spoIIG 이중포자변이주의 경우는 spoIIG 변이주와 더욱 닮은 것을 알 수 있었으며, 훨씬 주름진 것과 같은 투박한 세포벽 및 막을 가지고 있음을 관찰하였다. spoOJ 변이는 포자형성 빈도를 낮추고 aprE 활성을 감소시키는 반면, spoIIG 변이는 포자형성을 거의 하지 않으면서 aprE 활성에 상승효과를 가져왔다. spoOJ와 spoIIG 이중포자변이주는 spoOJ 변이의 효과는 거의 나타나지 않은채, spoIIG와 비슷한 aprE 활성을 보였다. Sporulation mutants of Bacillus subtilis were developed for overproduction of heterologous proteins. The strains spoOJ spoIIG, and spoOJ spoIIG double mutant were constructed from two pretense-delfted mutant (DB104). The vector containing aprE gene was integrated in the chromosome of each strain, then the morphology of each strain was observed by TEM (trasmission electron microscopy). The morphology of spoOJ mutant and spoIIG mutant coincides with the description of the previous reports, respectively. The sporulating cells of spoOJ SpoIIG double mutation resemble spoIIG mutant more similarly, but with a little rougher cell wall membrane. The spoOJ mutation in B. subtilis gives negative effect on aprE activity with only a decreased sporulation frequency. On the contrary spoIIG mutation increases the aprE activity twice with an undetectable sporulation frequency. In the case of spoOJ and spolIG, i. e. double mutation, the effect of spoOJ on aprE activity seems to be relieved and the double mutant shows more or less the same aprE activity compared to spoIIG mutant.
DNA Microarray Analysis of Immediate Response to EGF Treatment in Rat Schwannoma Cells
오민규,Daniel R. Scoles,Stefan-M. Pulst 한국생물공학회 2005 Biotechnology and Bioprocess Engineering Vol.10 No.5
Epidermal growth factor (EGF) activates many intracellular effector molecules, which subsequently in-fluence the expression levels of many genes involved in cell growth, apoptosis and signal transduction, etc. In this study, the early response of gene expressions due to EGF treatment was monitored using oligonucleotide DNA microarrays in rat schwannoma cell lines. An immunoblotting experiment showed the successful activation of EGF receptors and an effector protein, STAT5, due to EGF treatment. The microarray study showed that 35 genes were significantly induced and 2 were repressed within 60 min after the treatment. The list of induced genes included early growth response 1, suppressor of cytokine signaling 3, c-fos, interferon regulatory factor 1 and early growth response 2, etc. According to the microarray data, six of these were induced by more than 10-fold, and showed at least two different induc-tion patterns, indicating complicated regulatory mechanisms in the EGF signal transduction.
오민규,Sang-Woo Lee,박부수,Eun-Sil Choi 한국화학공학회 2010 Korean Journal of Chemical Engineering Vol.27 No.2
In our previous work, we developed an auxotrophic mutant of Saccharomyces cerevisiae sake kyokai No. 6strain for the selection of recombinant DNA. Sake yeast can accumulate high amount of S-adenosylmethionine (SAM)that is effective for treating depression, Alzheimer’s disease and so on. In this work, we further introduced SAM2 gene and ERC1, a frame-shift mutant of YHR032w, to maximize the production of S-adenosylmethionine in S. cerevisiae sake K6. The recombinant strain accumulated SAM about half of its dry cell weight
신정휴손(腎精虧損)으로 변증한 파킨슨병 환자의 이상운동증 치험(治驗) 1례(例)
오민규,김태용,김동진,신현수,Oh, Min-Kyu,Kim, Tae-Yong,Kim, Dong-Jin,Shin, Hyeon-Su 대한한방내과학회 2007 大韓韓方內科學會誌 Vol.28 No.4
Objectives : This study was designed to research the effects of oriental treatment on dyskinesia with lower limbs caused by Parkinson's Disease. Methods : We prescribed Yukmijihwang-tang (六味地黃湯) and Jaumganghwa-tang (滋陰降火湯) and treated acupuncture through Shinjungkyuk (腎正格) every day. It was used to treat the Shinjeonghyuson (腎精虧損) symptoms. We analyzed the frequency of dyskinesia over one night at intervals of three days and measured the score of 'Unified Parkinson's Disease Rating Scale' every week to evaluate the entire symptom's improvement. Results and Conclusion : He had been treated with western medicine to improve the dyskinesia symptoms, but couldn't get a noticeable result. Having oriental treatment for 43 days, dyskinesia in the lower limbs mostly disappeared and all symptoms of Parkinson's disease (tremor, rigidity, dysphagia, hallucination, anorexia, etc.) improved significantly.
Production of bio-based chemicals from Klebsiella pneumonia
오민규,정무영 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.0
The fermentative production of 2,3-butanediol has been intensively studied because of the potential of these products as platform chemicals, such as polymers, fuels, and plastics. Klebsiella pneumoniae is a good host for production of 2,3-butanediol from a wide range of carbon sources. We metabolically engineered K. pneumoniae (△wabG △ldhA △pflB) to reduce the production of byproducts. In particular, the inactivation of pflB significantly improved the yield of 2,3-butanediol from glucose to 92.2% of the theoretical maximum. We also engineered Enterobacter aerogenes for efficient utilization of sugarcane molasses in 2,3-butanediol production by elimination of carbon catabolite repression. Deletion of scrR and cra as well as overexpression of scrAB improved the strain’s performance of utilizing three sugars in sugarcane molasses and producing 2,3-butanediol.