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유기인계 및 카바메이트계 농약의 고감수성 아세틸콜린에스테라이즈의 대량생산
김영미,김소미,조문제,Kim, Young-Mee,K., Cho,Cho, Moon-Jae 한국응용생명화학회 2003 한국농화학회지 Vol.46 No.4
본 실험에서는 acetylcholinesterase(AChE, EC 3.1.1.7)를 이용한 간이 잔류농약 검사법에 필요한, 유기인계 및 카바메이트계 살충제에 대한 감수성이 증가된 AChE(MAChE)를 baculovirus를 이용하여 대량으로 생산하는 시스템을 구축하고 생산된 효소의 특성을 관찰하였다. 한라산에서 채취한 초파리에서 AChE의 cDNA를 합성한 후 PCR을 이용하여 AChE의 lipid anchor부분을 제거하고 site directed mutagenesis에 의해 E107Y, F368L, L408E의 염기서열을 변화시켜 재조합된 MAChE cDNA를 합성하였고 baculovirus vector에 삽입하여 대량생산을 시도하였다. 대량 증식에 필요한 조건으로 감염횟수가 네 번일 때, 그리고 세포수가 $2{\times}10^6$ cell/ml일 때 세포의 증식과 효소의 활성이 극대화됨을 알 수 있었다. His tag을 붙여 Ni-NTA affinity column을 이용하여 MAChE를 정제하였으며, 정제된 효소는 실험조건하에서는 pH(3-10)와 온도$(20-50^{\circ}C)$의 변화에 영향을 받지 않았다. 농약 추출액으로 methanol을 사용했을 때가 ethanol을 사용할 때 보다 효과적임을 알 수 있었다. 대표적인 유기인계와 카바메이트계 농약에 대한 저해율을 조사한 결과 재조합된 MAChE는 대만의 집파리 및 변형되지 않은 AChE에 비하여 전반적으로 농약에 대한 감수성이 높은 것으로 나타났다. For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells $(2{\times}10^6\;cell/ml)$ were infected four times at four-day-intervals. His-tag containing MAChE was purified using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures $(20-50^{\circ}C)$ under experimental conditions. As an extraction solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).
홍해삼 첨가 김치의 저장기간 중 품질 특성 및 기호도 변화
박수영 ( Soo Yeong Park ),임희경 ( Hee Kyoung Lim ),박상규 ( Sang Gyu Park ),조문제 ( Moon Jae Cho ) 한국응용생명화학회 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.2
To diversify usage of aquacultural red sea cucumber, new Kimchi containing red sea cucumber was developed and its quality and consumer preference were studied. The pH and total acidity, index of Kimchi ripening, decreased till 7 days of storage, then pH change became blunt and finally reached optimum pH 4- 4.5. The amount of reducing sugar gradually decreased during storage period. The number of lactic acid bacteria increased in early fermentation period in both Kimchi containing boiled and fresh red sea cucumber, however, more lactic acid bacteria found in fresh red sea cucumber containing Kimchi. Preference of Kimchi containing boiled or fresh red sea cucumber compared to control Kimchi containing fermented salted fish was evaluated. The results revealed there were no differences in color, flavor, texture, taste, after taste and overall preference.
간암세포주 Huh7에서 Hepatitis B virus X protein에 의한 간섬유화
손모아 ( Mo A Son ),박상규 ( Sang Gyu Park ),조문제 ( Moon Jae Cho ) 한국응용생명화학회 2016 Journal of Applied Biological Chemistry (J. Appl. Vol.59 No.1
Hepatitis B virus infection can cause hepatic fibrosis leading to cirrhosis and hepat℃ellular carcinoma. However the mechanism remains poorly understood. In this study, we found that Hepatitis B virus X-protein (HBx) increases vimentin, fibronectin, slug, snail and NOX4 expression. Because NOX4- mediated reactive oxygen species can increase slug and snail, which can induce fibrosis, HBx may be a key regulator of hepatic fibrosis development via NOX4 induction.
이정덕(Jeong Deuk Lee),이준영(Jun Young Lee),조백기(Baik Kee Cho),허원(Won Houh),심상인(Sang In Shim),조문제(Moon Jae Cho) 대한피부과학회 1989 대한피부과학회지 Vol.27 No.4
We report a case of congenital vellus hamartoma, which consists of numerous matured vellus hair follicles and sebaceous glands. This patient is 4-month-old male with multiple, variable sized nodules on the right epicanthsl fold area since birth. Histopathologically, some of the infundibular portion of the vellus hair shows irregular and reticulated hyperplasia. There are numerous well-formed vellus hair follicles surrounded with thick fibrous sheath and some of them are associated with rudiment sebaceous glands.
이준영(Jun Young Lee),박천욱(Chun Wook Park),조백기(Baik Kee Cho),허원(Won Hugh),조문제(Moon Jae Cho) 대한피부과학회 1986 대한피부과학회지 Vol.24 No.2
We report herein a case of subcutaneous granuloma annulare occured in a 3-year-old girl who had 7 subcutaneous nodules on the scalp, lumbar area, buttock, right tibial area and both ankles. liistopathologically, multiple foci of complete and incomplete collagen degeneration surrounded by histiocytes in a palisading arrangement were seen in the subcutaneous layer. Some vessel walls were thickened and infiltrated with inflammatory cells and many eosinophils were scattered among the inflamrnatory cell infiltrate around the palisaiing granuloma. Five of the lesions disappeared spontaneously and other s decreased in size without specific treatment in 2 months.
박홍진(Hong Jin Park),이동원(Dong Won Lee),최성우(Sung Woo Choi),조백기(Baik Kee Cho),조문제(Moon Jae Cho),심상인(Sang In Shim) 대한피부과학회 1995 대한피부과학회지 Vol.33 No.2
Carcinomatous degeneration of chronic scars, known as Marjcinulcer, are relatively rare. The majority of them are quamous cell carcinoma or other skin cancers including basal cell carcinoma. But malignant melanone, especially amelanotic melanoma oveloping from a chronic burn scar is extremely rare. We have experiencedl a case of amelanotic melanoma arising ron a chronic burn scar in a patient, 53 year-old female, who had a painful erythematous ulcra edonodule on the left heel for about 20 years since she had sustained a burn there at the age of 1 year. Although there are no melanin pigments in hematoxylinsin(H & E) stain as well a Fontana Masson stain, a biopsy specimen from the skin lesion revealed cytologic atypia and junctional activity in the dermoepidermal junction arcl positive immunohistochemical staier, for S-100 protein, which is consistent with amelanotic melanoma. lesion was widely excised and a skin graft was dure She was also treated with intralesional BCG injection and recurrence or metastasis was ot detected during the follow up period of 6 months. (Kor J Dermatol 1995;33(2): 339-343)
재래귤의 성숙시기별 리모노이드 쓴맛 표시자로서 Limonoid UDP-glucosyltransferase 발현 분석
김영미 ( Young Mee Kim ),이도승 ( Do Seung Lee ),전덕현 ( Deok Hyoen Jeon ),송연우 ( Yeon Woo Song ),이동선 ( Dong Sun Lee ),류기종 ( Key Zung Ryu ),조문제 ( Moon Jae Cho ),이동훈 ( Dong Hoon Lee ),김소미 ( So Mi Kim ) 한국식품저장유통학회(구 한국농산물저장유통학회) 2011 한국식품저장유통학회지 Vol.18 No.2
Limonoid UDP-glucosyltransferase (LUGT) is an enzyme that converts limonoids into their corresponding glucosides and ultimately ameliorates limonoid bitterness in Citrus species. In this paper, the LUGT gene was cloned via PCR from 10 Jeju Citrus species. All the deduced glucosyltransferase proteins harbored a highly conserved plant secondary product glucosyltransferase (PSPG) motif within the C terminal region. Phylogenetic analysis based on the amino acid sequence comparison of the LUGT proteins from 10 Citrus species generated three distinct types. The expression patterns of LUGT gene in three representative species from each type were quite different with that of C. unshiu Marc. cv. Miyagawawase(Gungcheon), which his without distinctive juice delayed bitterness. Ourresultssho wth at some Citrus speciessuchas Citrusleiocarpa HORT(Bingul), Citruserythrosa HORT (Dongjunggul), and Citrustachibana TANAKA(Honggul) end emicin Jeju maybe susceptible to intense juice delayed bitterness due to delay inexpression of LUGT.