Proteomics를 이용한 고랭지 배추의 고온장해 해석

신평균(Pyung-Gyun Shin),홍성창(Sung-Chang Hong),장안철(Ancheol Chang),김상효(Sang-Hyo Kim),이기상(Ki-Sang Lee) 한국생명과학회 2007 생명과학회지 Vol.17 No.12

무더운 날씨가 지속됨으로서 고랭지배추의 생장 및 결구가 지연되고 있는 강원도 정선군 질운산(새빗재)의 600 m와 900m의 배추를 사용하여 무기성분 및 단백질 발현패턴을 분석하였다. 식물체 무기성분에서는 생장에 관련된 질소 및 인산의 부족현상과 결구에 관련된 칼슘이 부족하였다. 단백체 분석은 2차원 전기영동에 의해 전체 126개의 단백질이 분리되었고 그 중 48개의 단백질이 고도에 따라 변화하는 양상을 보여주었다. 이 중에서 30개의 단백질 서열이 결정되었는데, 해발 900 m에서 단백질 발현이 증가한 14개 중에서 oxygen- evolving proteins, rubisco activase and ATPase 등이, 해발 600 m에서는 glutathione S-transferase (1, 28 kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein 등 16개의 단백질 발현이 증가하였다. 이러한 단백질은 식물체 손상에 대한 보호기작을 가진 스트레스관련 단백질로 가뭄, 온도상승, 밤낮의 온도차 등의 반복으로 복합적이며 동시 다발적으로 나타나는 고온장해 현상으로 사료된다. High temperature stresses have caused growth inhibition and delayed heading in highland cultivation Chinese cabbage during summer in Korea. We have studied high temperature stress responses in the terms of changes of inorganic components and proteins by proteomic analyses. Insufficiencies of nitrogen and phosphorus have affected growth rate and calcium deficiency has caused blunted heading. Proteins extracted from Brassica seedling grown at the altitude of 600m and 900m in the Mount Jilun were extracted and analysed by 2-dimentional polyacrylamide gel electrophoresis. Profiles of protein expression was then analyzed by 2-dimentional gel analyses. Protein spots showing different expression level were picked using the spot handling workstation and subjected to MALDI-TOF MS. Total 48 protein spots were analyzed by MALDI-TOF MS and 30 proteins spots out of 48 were identified by peptide mass fingerprinting analyses. Fourteen proteins were up-regulated in extracts from the altitude of 900m and they were identified as oxygen-evolving proteins, rubisco activase and ATPase etc. Sixteen proteins were up-regulated in extracts from the altitude of 600m and they were identified as glutathione S-transferase(1, 28 kD cold induced- and 24 kD auxin-binding proteins) and salt-stress induced protein etc. These stress-induced proteins were related to the mediated protective mechanism against oxidative damage during various stresses. The results indicated that physiological phenomenon in response to high temperature stresses might be resulted by complex and multiple array of responses with drought, heat, oxidative, salt, and cold by high temperature.

수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정

신평균(Pyung-Gyun Shin),장안철(Ancheol Chang),홍성창(Sung-Chang Hong),이기상(Ki-Sang Lee) 한국생명과학회 2007 생명과학회지 Vol.17 No.12

BAK1(Brassinolide insensitive associated receptor kinase 1)는 브라시노스테로이드 생합성 대사관련 신호전달 매체이다. BR 생합성 및 신호전달 돌연변이체는 매우 특징적인 난쟁이 표현형을 보인다. 과채류전용 양액배지인 Sonneveld 양액을 이용하여 양분결핍에 의해 왜성을 나타내는 토마토를 선발하였다. 선발된 토마토에 대해 이차원 전기영동법으로 단백질체를 분석한 결과, 발현차를 나타내는 28개의 단백질 spot이 분리되었다. 분리된 단백질 spot 중 현저하게 발현이 억제된 단백질 spot 6개를 선발하여 단백질 서열을 결정하였다. 실험결과, pI 4.5, 분자량 24 kDa를 나타내는 단백질은 브라시노스테로이드 생합성에 관여하며 왜성 표현형을 나타내는 신호전달 단백질, BAK1으로 동정되었다. BCK1, cystein proteinase, sulfutase, peroxidase, zinc finger factor로 동정된 나머지 단백질들은 브레시노스테로이드 생합성관련 신호전달기작에 관여하는 단백질로 추정되었다. BAK1을 검정하기 위해 단백질 서열이 결정된 부위로부터 프라이머를 디자인하여 RT-PCR를 수행한 결과, 증폭된 500 bp의 산물이 정상과 발현차를 보여주었는데 이 결과는 양분조절에 의해서도 BAK1의 발현이 조절될 수 있음을 시사한다. Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

데이터 분석을 통한 도시가스 사용량 예측 프로세스 개발

신평식(Pyung Sik Shin),주경민(Kyung Min Ju),이정우(Jeong Woo Lee) 한국가스학회 2019 한국가스학회 학술대회논문집 Vol.2019 No.5

팔레높시스의 고랭지재배 및 교잡육종

신평균(Shin Pyung-Gyun),류승열(Ryu Seung-Yeul),유동림(You Dong-Lim),남춘우(Nam Chun-Woo),조우석(Cho Woo-Suk) 한국원예학회 1998 원예과학기술지 Vol.16 No.3

연구단보 : 건조 및 저장조건에 따른 쌀 저장단백질의 변화양상

신평균 ( Pyung Gyun Shin ),장안철 ( An Cheol Chang ),홍성창 ( Seong Chang Hong ),이기상 ( Ki Sang Lee ),이금희 ( Keum Hee Lee ),이용복 ( Yong Bok Lee ) 한국환경농학회 2008 한국환경농학회지 Vol.27 No.4

Quality of rice grain changes during dry storage with internal physiological changes and external injury by organism. Storage rice changes by condition with respiration via variable temperature, hydrolysis enzyme reaction, lipid peroxidation occurs with change of palatability. During dry storage, physiological change with protein variation pattern was examined by image analysis on proteomic technology. Analysis revealed that protein activity had no change store at room temperature and store at 40℃, but decreased store at 60℃. Analysis of variable hydrophobic protein pattern revealed that protein activity of beta-tubulin, protein disulfide isomerase, vacuolar ATPase b subunit, globulin was not significantly decreased all dry and store condition. However, heat shock protein 70, and glutathione transferase was significantly decreased when rice dried at 60℃ compared with room temperature and 40℃ dry condition.

In Vitro Translation of Nodule Specific mRNA from Alfalfa (Medicago sativa L. Vernal) Root Nodules

신평균,김경수,조무제,Shin, Pyung-Gyun,Kim, Kyung-Sao,Cho, Moo-Je 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.2

알팔파근류에 R. meliloti 102F51 및 3A150 mutant ($nif^-$, $nod^+$)를 접종하여 얻은 nodule과 접종하지 않은 뿌리로부터 $poly(A)^+-mRN$를 분리 정제하여 [$^3H$]-leucine 및 [$^{35}S$]-methionine으로 in vitro translation 시킨 후 이들을 leghemoglobin 및 nodule specific protein antibody를 이용하여 immunoprecipitation 시킨다음 2D-polyacrylamide gel 전기영동을 행한 결과 분자량 12,000-30,000 범위내에서 적어도 25종 이상의 근류에서 선택적으로 발현이 촉진되는 단백질들을 확인하였으며 이들은 대두의 경우와 상당한 차이가 있는 것으로 보아 두 과작물간에 이들 단백질 양상이 서로 차이가 있을 것으로 추정된다. Nodule specific proteins (nodulins) were detected by the 2D-polyacrylamide gel electrophoresis of the immunoprecipitated in vitro translation products of the $poly(A)^+-mRN$ A from alfalfa root nodules. At least 25 nodule specific polypeptides in the molecular weight range of 12,000-30,000 were identified in the Rhizobium meliloti 102F51 (wild type) infected alfalfa root nodules. The nodulins accounted for about 40% of [$^3H$]-leucine labeled and 12% of [$^{35}S$]-methionine labeled proteins.

Gliocladium virens 와 Trichoderma harzianum 의 속간 원형질체융합

신평균,조무제 ( Pyung Gyun Shin,Moo Je Cho ) 한국균학회 1993 韓國菌學會誌 Vol.21 No.4

느타리버섯 소포자 형성 균주 개발

신평균,오세종,유영복,Shin, Pyung-Gyun,Oh, Se-Jong,Yoo, Young-Bok 한국버섯학회 2006 한국버섯학회지 Vol.4 No.2

느타리버섯의 포자는 버섯재배 농업인의 호흡기계통질환인 알레르기를 유발한다. 느타리버섯의 무포자형성 균주는 재배농업인의 건강에 도움을 줄뿐 아니라 다른 재배사에 바이러스병 감염의 위험을 감소시킬 수 있다. 소포자 형성 느타리버섯을 육성하기 위해 무포자형성 균주의 선발 및 특성검정 결과 ASI 2069 균주가 무포자이면서 수량성이 높으나 형태적으로 상품적 가치가 없어 원형질체 재생에 의한 단핵화를 시도하여 neohaplont를 분리하였다. 원형 1호(ASI 2180) 및 무포자 느타리간의 단핵교배 (Mon-Mon)를 시도한 결과 128교잡체를 얻어 이 중 30균주를 특성 검정한 다음 자실체형성 및 담자 포자비산량 조사에 의해 소포자형성 유망 13균주를 선발하였다. 그 중에서 G192 교잡체가 소포자이면서 수량성이 좋은 가장 우량 균주로 선발되었다. The enormous production of spores by the fruitbodies in the cultivation of oyster mushrooms (Pleurotus ostreatus) is develop an allergy with symptoms similar to an "extrinsic allergic alveolitis (EEA)". the sporeless strain would noy only benefit health of mushroom workers but also reduce the risk of viral infections on the mushroom farms. For the development of a sporeless strain of P. ostreatus we used strain ASI 2069. This non-commercial strain is completely nonsporulating. We have recovered both nuclear types of strain ASI 2069 as monokaryons (hereafter referred to as neohaplonts) by protoplasting the mycelium. Crosses between neohaplonts and SSI's(single spore isolates) obtained from a sporulating commercial strain ASI 2180 yielded fruitbodies that isolated 128 strains. 13 excellent strains are selected from 30 bred strains by quality of fruitbodies and spore number. Among 13 excellent strains, G192 strain is chose finally to high yield and sporelessness.

Gliocladium virens 와 Trichoderma harzianum 의 속간 핵 전이체의 효율적 선발

신평균,유영복,류진창,박용환,조무제 ( Pyung Gyun Shin,Young Bok Yoo,Jin Chang Ryu,Young Hwan Park,Moo Je Cho ) 한국균학회 1994 韓國菌學會誌 Vol.22 No.3

도라지의 약리성분

정진환,신평균,류진창,장대식,조성환 ( Jin Hwan Chung,Pyung Gyun Shin,Jin Chang Ryu,Dae Sik Jang,Sung Hwan Cho ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.2

Platycodon root contains abundant pharmaceutical substances and is widely used as a food and a medicinal herb. The three-year-old and the 24-year-old Platycodon roots cultivated in a local farm were analyzed and compared with their pharmaceutical substances to identify the major components. Reverse column HPLC analysis of ethanol extracts from the 24-year-old roots showed some distintive peaks after the retention time of 50 min which were absent in the extracts from the three-year-old roots. This indicates that several pharmaceutical substances are present in the older roots. There were no differences in sterol composition and saponin content between the 24-year and three-year-old roots. The 15-year-old and the 23-year-old Platycodon roots, the 6-year-old Ginseng root, and the three-year-old Condonopsis root all did not show significant anticomplement activity.