국내에서 분리된 Bacillus thuringiensis NT0423은 나비목과 파리목 곤충에 독성을 보이는 130 kDa의 전형적인 다이아몬드형 내독소 단백질을 생성한다. 이 균주의 파리목에 대한 독성을 강화하기 위하여, B. thuringeinsis NT0423에 모기 유충에 강한 독성을 보이는 B. thuringiensis subsp. morrisoni PG-14의 cryVID유전자를 가지고 있는 pCG10 플라스미드를 electroporation 방법을 이용하여 형질전환하였다. 형질전환체인 B. thuringiensis PT1227내에서 cryIVD와 숙주가 생성하는 원래의 다이아몬드형 130kDa 내독소 단백질 유전자는 그 자신의 형태로 잘 발현되었다. 형질전환체의 모기 유충에 대한 독성은 원래 숙주의 내 독소 단백질과ㅏ 도입된 CryIVD의 상승효과에 의해 현저히 증가하였다.

정확한 in vitro 전사가 일어날 수 있는 진딧물의 세포추출액을 제조하였다. 전사를 직접 조절할 수 있는 단백질 인자를 규명하기 위하여 전사개시점과 그의 상류에 결합하는 DNA 결합단백질을 탐색했다. 전사개시점을 포함하는 단편 A(-194/23)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합했으며 전사개시점 상류의 DNA 단편 B(-393/-263)에는 52kDa, 50kDa, 40kDa의 단백질들이 결합한 반면 DNA 단편 C(-263/-195)는 53kDa단백질만이 결합했다. 그리고 이들 DNA 결합단백질들의 DNA 결합 활성에는 양이온이 요구되었다.

Bacillus thuringiensis kurstaki와 aizawai의 내독소단백질을 알카리용액 또는 트립신, 곤충소화액 드응ㄹ 처리하여 전기영동한 후 단백질팬턴을 비교하였다. 두 균주의 주요 결정단백질은 130kd와 64kd의 단백질이었으며, 소화액이나 효소로 처리한 경우 공통적으로 62kd의 활성독소가 생성되었다. 그러나, aizawai는 kurstaki에 비해 현저히 적은 양의 62kd 단백질을 생성하였다. 흰불나방의 유충이 Bacillus thuringiensis 독소를 섭식하였을 때 지방체를 비롯한 몇 가지 조직에서 45kd의 스트레스 단백질이 유발되었는데 이 단백질은 열충격이나 저온 충격시에도 마찬가지로 생성되었다.

멧누에나방(Bombyx mandarina) 난각유전자의 형질발현기구를 규명하기 위한 연구의 일환으로 각형성과정에 있어서 난각단백질의 합성과정을 분석하였다. 난각단백질의 합성과정을 여포세포의 기난관배양에 의한 in vitro합성계에 의하여 분석한 결과, 합성과정은 8단계로 나눌 수 있었으며 SDS-PAGE 분석에 의해 17개 이상의 단백질 band로 분리되었다. 멧누에 난각단백질은 합성시기에 따라 3단계로 구분되며 초기에는 비교적 고분자 성분이, 합성의 중후기에는 비교적 저분자성분이 합성되는 등 난각단백질의 합성은 시기특이적으로 조절되었다.

To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03-BMOC-3 medium at 37 for 96 hrs. under the atmosphere of 5% and 95% air. Fibronectin, gelatin, or collagen significantly(P1.4, 45.4i1.4, and 44.8O.9, respectively. And the diameter of those eggs ranged 104.61.9, 102.82.3, and 103.4O.8 m, respectively.

본 연구(硏究)는 식물(植物)의 발생(發生)과 분화(分化)에 관(關)한 생화학적(生化學的) 분자생물학적(分子生物學的) 구명(究明)을 위해 조직(組織) 발생(發生) 단계(段階)의 특이적인 유전자(遺傳子)의 발현조절 기작과 역할(役割)을 연구(硏究)하기 위한 기초(基礎) 지식을 얻고자 수행(遂行)되었다. 벼를 실험재료(實險材料)로 하여 현미로부터 callus를 유도(誘道)하고 이들 callus에서 여러가지 방법(方法)을 통(通)해 embryogenic callus와 nonembryogenic callus를 분리(分離)해 somatic embryogenesis에 관련된 특수한 단백질(蛋白質)을 찾고자 하였고 그 결과(結果)를 요약(要約)하면 다음과 같다. 지금까지의 보고(報告)에 의하면 재분화가 잘되는 callus와 그렇지 않은 callus간에는 여러가지 면에서 차이가 난다고 알려져 있다. 재분화가 잘되는 callus는 표면이 거칠고 흰색에 가까우며 조직(組織)이 단단하다. 반면에 재분화가 되지않는 callus는 표면(表面)에 윤기가 나며 단단하지 않다. 이러한 보고(報告)를 기초(基礎)로 하여 media에 1mg/1의 2,4-D가 첨가(添加)된 배지(培地)에서 2주 정도 callus를 유기시키고 유기된 callus는 다시 배양(培養)하였다. 7-8차 계대배양하면서 embryogenic callus와 nonembryogenic callus를 분리(分離)한 다음 이를 시료(試料)로 하여 protein을 추출(抽出)하여 SDS-PAGE 상에서 비교하여 보았다. 그 결과(結果) 두 callus간에 차이가 있음을 알 수 있었다. 7-8차 계대배양한 두 종류(種類)의 callus로 부터 protein을 분리하여 two-dimensional gel 전기영동에 의해 비교하여 본 결과(結果) 몇 개의 단백질(蛋白質)이 서로 다름을 알 수 있었다.

배추흰나비 P. rapae와 P. brassicae GV에 대한 봉입체 및 바이러스단백질의 물리화학적 성상을 비교 분석한 결과 전기영동법에 의한 봉입체단백질은 그 분자량이 P. rapae GV는 30kd, P. brassicae GV는 31 kb의 단일단백질이었으며, trypsin, chymotrypsin, papain 및 staphylococcus aureus V8 protease에 의한 peptide mapping에서는 두 바이러스 간에 큰 차이가 없었다. 바이러스입자단백지르이 전기영동법에 의한 분석결과, 두 바이러스 간에 큰 차이가 없었다. 바이러스입자단백질의 전기영동법에 의한 분석결과, 두 바이러스에서 각각 42개의 band가 나타났으며 고분자량 부분에서 P. rapae GV의 경우는 115, 110, 105 및 103 kd의 4개 band인데 반해 P. brassicae GV는 107 kd의 한 band만 관찰되었다. 그리고 봉입체단백질의 아미노산 분석비교에서 두 바이러스 간의 큰 차이는 없었으며 일반적으로 곤충에 많이 존재하는 산성아미노산인 aspartic acid와 glutamic acid의 함량이 많았고 염기성 아미노산에서 lysine의 함량이 특히 많았으며 봉입체단백질의 면역학적인 시험결과에서는 P. rapae GV 항혈청에 대해 두 바이러스는 공통 침강선을 형성하였다.

담배거세미나방(Spodoptera litura: SI), 누에(Bombyx mori: Bm) 및 흰불나방(Hyphantria cunea : Hc)으로부터 분리된 핵다각체병 바이러스(nuclear polyhedrosis virus : NPV)의 다각체의 단백질의 특성과 그에 대한 alkaline protease의 영향을 SDS-PAGE 및 혈정학적 방법으로 분석했다. Alkaline protease를 부활화시킨 후 다가체 단백질을 SDS-PAGE한 결과, BmNPV의 경우 30kD, SINPV와 HcNPV는 31kD의 단일 major band 및 이것들의 종합체(polymer)인 57kD와 66kD의 minor band들이 관찰되었다. Alkaline protease의 부활화를 성략한 SI NPV 다각체를 알칼리 용액으로 시간 차이를 두고 처리한 후 SDS-PAGE한 결고, 알칼리 처리시간이 경과함에 따라서 alkaline protease활성에 의해 다각체 단백질이 일정한 pattern으로 저분자화됨이 뚜렷하였다. SINPV 와 BmNPV 다각체 단백질의 항체를 제조하여 SINPV, BmNPV및 HcNPV 다각체 단백질간의 혈정학적 상동성을 이중형역광산법과 Western blot으로 비교한 결과는 3종 모두에서 공통 antigenic determinants의 존재가 인정되었으며 major 다각체 단백질의 종합체 형성과 alkaline protease에 의한 분해를 확인했다.

흰불나방 (Hyphantria cunea Drury) 의 용휴면이 결정되는 시기, 휴면용에서의 20-hydroxyecdysone 의 영향과 전기영동 방법으로 측정된 용단백질의 변화는 다음과 같았다. 용휴면의 결정은 유충발육기간중 초기 5~15일에 이루어졌다. 산소소비율의 변화는 정상발육용에서 U자 모양의 곡선을 나타내었고, 휴면용에서는 l TEXO2/TEX /g/hr 수준으로 감소되어 유지되었다. 휴면용에서 추출된 2.5%균질액은 20-hydroxyecdysone 의 처리로 호흡활성이 증가하였고, 휴면예정용은 직접 주사로 우화하였다. 수용성 단백질의 함량 변화는 정상발육용에 급격하게 이루어졌으나, 휴면용에서는 완만하였고, 더 많은 양이 포함되어 있었다. 휴면예정용의 지방체에서 분리한 단백질의 종류는 비휴면용의 경우와 달랐다.

S100 protein family is small calcium-binding proteins with two EF-hand motifs and comprises more than 20 proteins in human. Although S100A proteins are known to play important roles in proinflammatory responses including damage-associated molecular pattern (DAMP) signaling and in the establishment of pregnancy, the expression of S100As have not been determined in the uterine endometrium during the estrous cycle in pigs. Thus, this study was performed to investigate expression and localization of S100A8, S100A9, and S100A12 in the uterine endometrial tissues during the estrous cycle in pigs. Real-time RT-PCR analysis showed that S100A8, S100A9, and S100A12 mRNAs were expressed in the uterine endometrium during the estrous cycle with higher levels on days 15 and 18 of the estrous cycle than the other days of cycle. Immunohistochemistry analysis showed that S100A9 and S100A12 proteins were mainly localized to the immune cells in the uterine endometrium. Especially, S100A9- and S100A12-positive immune cells were detected in the uterine blood vessels on day 15 of the estrous cycle, and also localized to stroma near to luminal epithelium on days 0 and 18 of the estrous cycle. These results showed that S100As were expressed in the uterine endometrium during the estrous cycle in a cyclic stage-specific manner, and these proteins were localized to the immune cells in the endometrium. These suggest that immune cells expressing S100A proteins may be recruited into the endometrium during the estrous cycle and play an important role in regulating endometrial function in pigs.

This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta (Sec61β), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVMⅠ) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVMⅡ). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, Sec61β, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVMⅠ or IVMⅡ stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, Sec61β and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but Sec61β and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, Sec61β, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of Sec61β and COPG2 could be changed by EGF in the porcine oocytes during maturation.

Background: Prenatal exposure to infectious and/or inflammatory insults can increase the risk of developing neuropsychiatric disorder such as bipolar disorder, autism, and schizophrenia later in life. We investigated whether Valeriana fauriei (VF) treatment alleviates prepulse inhibition (PPI) deficits and social interaction impairment induced by maternal immune activation (MIA).Methods and Results: Pregnant mice were exposed to polyriboinosinic-polyribocytidilic acid (5㎎/㎏, viral infection mimic) on gestational day 9. The adolescent offspring received daily oral treatment with VF (100㎎/㎏) and injections of clozapine (5㎎/㎏) for 30 days starting on the postnatal day 35. The effects of VF extract treatment on behavioral activity impairment and protein expression were investigated using the PPI analysis, forced swim test (FST), open field test (OFT), social interaction test (SIT), and immunohistochemistry. The MIA-induced offspring showed deficits in the PPI, FST, OFT, and SIT compared to their non MIAinduced counterparts. Treatment with the VF extract significantly recovered the sensorimotor gating deficits and partially recovered the aggressive behavior observed in the SIT. The VF extract also reversed the downregulation of protein expression induced by MIA in the medial prefrontal cortex.Conclusions: Our results provide initial evidence of the fact that the VF extract could reverse MIA-induced behavioral impairment and prevent neurodevelopmental disorders such as schizophrenia.

Dynamic reorganization of actin filaments is essential for various stages of mammalian oocyte maturation, including spindle migration, actin cap formation, polar body extrusion, and cytokinesis. Various actin binding proteins (ABPs) have been known to be involved in the regulation of actin filament remodeling. We elucidate roles of three different actin binding proteins in mouse oocyte maturation. The heterodimeric actin-capping protein (CP) binds to the fast-growing(barbed) ends of actin filaments and plays essential roles in various actin-mediated cellular processes. When CP is knockdowned or inhibitory component was overexpressed, asymmetric division of oocyte have been compromised. It turns out that knockdown or inhibition of CP deplete cytoplasmic actin mesh level, which have been known to be essential for maintain cytoplasmic actin mesh. Another actin binding proteins, tropomodulin 3 (Tmod3), binds to the slow-growing end of actin filaments and knockdown or expression deletion mutant of Tmod3 also decrease actin mesh level in maturing oocyte and it severely ablated asymmetric division of oocyte. Finally, tropomyosin 3, actin filament binding proteins protect actin filament from depolymerization, is also important to maintain cortex integrity in maturing oocyte. Taken together, these finding showed the essential roles of actin binding proteins in remodeling of actin filaments in mammalian oocyte development.

Understanding the response of a crop to water deficiency is the first step towards breeding drought-tolerant varieties. In this study, inbred maize (Zea mays L.) lines KS140 and KS141 were subjected to drought stress by withholding water for 10 days at the V5 or V6 leaf stage. Water-deficient plants experienced a decrease in relative leaf water content, stomatal conductance, net CO2 assimilation rate, and water use efficiency compared to well-watered plants. This was accompanied by a decrease in the relative leaf water content that resulted in severe growth retardation in KS140 and KS141. However, leaf chlorophyll content in KS140 was unchanged. To understand the proteome dynamics during the 10-day drought stress in maize leaves, comparative proteome analysis was carried out between the well-watered and water-withheld leaves. Differential expression was observed for 29 protein spots from KS140 and 14 protein spots from KS141, and these were identified using MALDI-TOF mass spectrometry. Among identified proteins, metabolism and stress related proteins were highly were increased by drought stress. This study provides a protein profile of a Korean maize inbred line during drought stress, which will be valuable for future studies of the molecular mechanisms underlying drought resistance and for development of selective breeding markers for drought tolerance in maize.

Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (≥ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose- 1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.