곤충 유전자를 이용한 항세균성 펩타이드 생산 및 응용에 관한 기초 연구로서 비병원성 세균(Escherichia coli K12)으로 면역 반응을 유도한 누에로부터 발현량이 증가하는 유전자 중 Hyalophora cecropia의 attacin과 cDNA 상동성을 나타내는 BmInc6 클론을 분리하고 그 특성을 조사하였다. BmInc6 cDNA의 전체염기서열을 분석한 결과 그 크기는 852 bp이고, 35번째 염기에서 변역이 개시되어 679 bp 위치에서 종결되는 open reading frame을 가지며 812번째 위치에 잠정 전사 종결 신호의 존재가 확인되었다(GenBank, AF005384). BmInc6에 의해 coding되는 아미노산은 214개이며, hydropathy 분석 결과 친수성을 나타내는 단백질이었다. 그리고 BmInc6 유전자에 의해 연역되는 펩타이드를 nuecin으로 명명하였다. Nuecin 유전자를 baculovirus 발현 백터계를 이용하여 곤충세포주에서 발현시킨 결과 전사체의 크기는 약 950 bp였고, 세포내 벌현 펩타이드의 분자량은 약 23 kDa이었다. 세포내에서 발현된 nuecin 전구체로 추정되는 23 kDa 펩타이드는 세포외로 분비되는 과정에서 약 3 kDa의 signal 펩타이드가 제거됨으로서 약 20 kDa의 성숙 nuecin으로 된다는 사실을 단백질 전기영동상으로 확인하였다. Nuecin 단백질의 항세균 활성을 수종의 그람 음성 및 양성 세균에 대해 검정한 결과, 특히 E. coli와 Bacillus subtilis에 높은 활성을 나타내었으며, attacin이 한정된 그람 음성 세균에만 항세균 활성을 나타내는데 비해 nuecin은 그람 음성은 물론 그람 양성에도 항세균 활성을 나타내어, 보다 넓은 항세균 스펙트럼을 가지는 것을 알 수 있었다.있는 자격이 주어지므로 대부분의 보조적 보장소득 수혜 노인들은 단순히 금전적인 수입의 측면뿐 아니라 그 외의 사회보장 프로그램에 의존하고 있다. 이러한 면에서 보조적 보장소득의 수혜는 한국 영주권자 노인들의 독립적인 삶에 큰 역할을 하고 있다. 이러한 사회복지 프로그램을 영주권자들에게는 전면 금지하는 새로운 사회복지법은 재미 한국 영주권자 노인들뿐 아니라 그들의 가족들에게도 막대한 경제적 부담을 줄 것이다. 본 논문은 영주권자 노인들을 대상으로 분석한 것이지만, 주거형태를 비롯한 일반적인 특성을 비교해 보면 영주권자, 귀화 시민권자, 그리고 미국태생 노인들은 다른 성격의 집단들임을 보여주고 있다. 종래의 소수민족 노인들에 관한 연구들이 이민자 노인들을 영주권자와 귀화 시민권자의 구분없이 하나의 집단으로 간주하고 분석해 왔던 것을 볼 때, 앞으로의 연구는 이론적으로나 방법론적으로 시민권의 유무가 주거형태에 끼치는 영향도 함께 고려해야 할 것이다.에 나타난 인도의 영향은 여성복식과 남성복식에 있어서 서로 유사점과 차이점이 보이는데, 인도의 영향이 여성복식에 있어서 그 빈도가 더 높고, 종류가 더 다양함을 볼 수 있다. 여성복식에 있어서는 12가지의 다양한 인도복식스타일이 나타났으며, 그중 가장 많이 보이는 스타일은 Indian Shirt/Blouse/Smock/ Dress이며, 그 뒤를 이어 Madras, Indian lowery등을 볼 수 있다. 남성복식애 나타난 7가지의 스타일 중에는 Madras가 가장 빈도가 높으며 그외의 스타일들은 그 빈도가 매우 낮음을 볼 수 있다. 인도의 영향의 정도 (Attribution Categories) 있어서는 여성과 남성복식 모두에 있어서 인도에서 직접 수입된(originated) item이 각각 전체의 90%와 81%를 차지하여, 인도복식의 영향은 받았으나

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.

Bacillus thuringiensis subsp. kurstaki HD-1으로부터 생산된 살충성 내독소 단백질을 coding하는 CryIIA 유전자를 클로닝하고 염기서열을 조사하였다. HD-1 균주의 12개 plasmid 중 225kb plasmid를 분리하여 CryIIA 유전자를 포함하는 5kb HindIII 절편을 hybridization하여 찾아냈다. 이 절편을 plasmid pUC19에 ligation하여 E. coli에 형질 전환하였다. 이 독소 유전자를 포함하는 4kb BamHI-HindIII 절편은 vector pT7-5에 ligation하여 pSKIIA라하였다. pSKIIA는 3개의 open reading frames(orf1, orf2, orf3)로 구성되어 있으며 염기서열은 3,952base로 되어 있었다. 이러한 3개의 orf 각각의 발현 여부를 확인하기 위하여 생물검정을 하였다. 그러나 orf1 또는 orf2에 의한 형질 전환체는 독성이 없는 것으로 나타났다. orf3를 포함하는 형질 전환체는 3종의 나비목 곤충(배추점나방, 담배거세미나방, 담배나방) 및 1종의 파리목 곤충(집모기) 유충에 대하여 독성을 나타내었다.

한국산 멧누에(Bomdyx mandarina)chorion 유전자의 형질 발현기구를 규명하기 위한 연구의 일환으로 본 실험을 실시했다. 멧누에의 난각구조를 주사형 전자현마경에 의해 관찰한 결과 매우 특이적인 구조가 인정되었다. 즉 원추상의 불규칙 돌기에 의한 돌기구조층과 이 돌기 구조층을 덮고 있는 한 층의 얇은 덮개 구조층이 그것이다. 2차원 전기영동법에 의해 chorion 단백질을 분석한 결과, 난각을 구성하는 주요 단백질 성분은 등전점 4~6, 분자량 6~30 kd로 밝혀졌다. 특히 특이난각구조와 관련된 특이단백질 성분을 검출하였으며 이들의 대부분은 고cysteine 단백질인 것으로 추정했다. 이상의 연구결과에 의해 멧누에 난각의 특이 구조 형성에 따른 유전자발현기구 규명을 위한 기초자료가 얻어졌다.

Water temperature is one of the most important factors of fish survival, affecting the habitat, migration route, development, and reproduction. This experiment studied the induction level of heat shock protein (HSP70) mRNA and protein in a walleye pollock (Gadus chalcogrammus) primary hepatocyte culture based on different temperatures. Hepatocytes were attached at 7.5 °C for 24 hours. Hsp70 induction levels were then measured for 48 hours at 5, 8, 11, 14, and 17 °C. The induction level was lowest at 5 °C and generally increased with temperature until 14 °C. The induction level was reduced at 17 °C, indicating that 14 °C is the highest tolerable temperature for hepatocytes. These data indicate that primary hepatocyte cell culture is under no stress at 5 and 8 °C. Temperatures greater than 11 °C induce stress, showing similar induction patterns in both mRNA and protein in hepatocytes. The results suggest that 14 °C is the maximum internal defense temperature of walleye pollock survival.

CREBZF (cAMP-response element binding protein zhangfei) is a member of ATF/CREB family, and which regulates various cellular functions by suppressing major factors with direct interaction. In this study, we have examined the expression of CREBZF on mouse endometrium during uterus estrous cycles and estrogen (E2) treatment. In uterus, CREBZF mRNA expression was higher than other organs and mRNA and protein of CREBZF was increased in proestrus phase and decreased in estrus phase. The expression of CREBZF in 3-weeks old mouse uterus was reduced by E2 injection in endometrium. In addition, the expression of progesterone receptor, a marker of E2 in ovariectomized mice was found to be strongly expressed in stroma, while CREBZF was only expressed in epithelium. Also, we conformed that E2-suppressed CREBZF was restored by co-injection of ICI 182,780, an estrogen receptor antagonist. Overall, these results suggest that CREBZF is regulated by estrogen and involved in ER signaling pathway in mouse uterus.

Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

The germline stem cells of the Drosophila ovary continuously produce eggs throughout the life- span. Intricate regulation of stemness and differentiation is critical to this continuous production. The translational regulator Nos is an intrinsic factor that is required for maintenance of stemness in germline stem cells. Nos expression is reduced in differentiating cells at the post-transcriptional level by diverse translational regulators. However, molecular mechanisms underlying Nos repression are not completely understood. Through three distinct protein-protein interaction experiments, we identified specific molecular interactions between translational regulators involved in Nos repression. Our findings suggest a model in which protein complexes assemble on the 3’ untranslated region of Nos mRNA in order to regulate Nos expression at the posttranscriptional level.

골 형성 단백질(Bone morphogenetic protein, BMP)은 TGF-β superfamily의 구성원 중에 하나이며, 이 들은 원래 뼈 형성을 유도하는 능력에 의해 발견되었지만, 곧 세포분열, 세포자살, 세포이동 그리고 세포 분화를 조절하는 것으로 밝혀졌다. 매우 다기능적인 골 형성 단백질은 포유동물의 reproduction에도 매우 중요한 역할을 하지만 알려진 바가 적다. 그래서 본 연구에서는 골 형성 단백질이 착상 전 oviductal environment에 어떠한 영향을 미치는지 밝히고자 하였다. 먼저, Estrus 시기 생쥐의 oviduct cDNA를 통해 RT-PCR한 결과, BMP1, 2, 3, 4, 5, 6, 7, 8, 10, 15 mRNA와 BMP receptor인 BMPR1A, BMPR1B, BMPR2 mRNA의 발현을 확인하였고, BMP Inhibitor인 Noggin, Follistatin, Chordin mRNA의 발현 또한 확인하였다. 이들 중 BMP2, 3, 7, 15 mRNA의 level이 BMP5, 6, 10 mRNA의 level보다 많은 발현량을 보였다. BMP receptor 중에서는 BMPR2 mRNA의 level이 제일 높았다. BMP Inhibitor 중에서는 Chordin mRNA의 발현량이 제일 많았다. 이들 가운데 가장 높은 발현 수준을 보인 BMP2의 단백질의 localization은 Immunohistochemistry를 통해 확인하였다. BMP receptor와 BMP Inhibitor에 대한 연구는 진행 중에 있지만, 생식주기별 생쥐의 oviduct 내에서 BMP2 단백질의 발현수준은 proestrous, metestrous, diestrous보다 estrous stage에서 가장 강한 발현을 보였다. 이는 BMP2가 steroidogenesis에 영향을 주어 oviductal cell proliferation에 관여한다는 것을 보여준다.

Soybean is very useful crop to supply vegetable protein for human. However, cultivation arear of this economically important crop is gradually diminished in upland field. Hence, cultivation area of soybean is increased in paddy field. During the growth duration of soybean, excessive moisture injury is serious problem for sustainable production and supply. We investigated protein expression according to different period of seed soaking and germination after seed soaking. For comparison on expression of protein according to different condition, we performed two-dimensional electrophoresis. After electrophoresis analysis, we selected differentially expressed protein spots according to different condition such as soaking period and germination after soaking to identify protein function by using MALDI-TOF. Results revealed that pattern of expression of protein according to soaking period and germination after soaking were generally not different in major spots. However, degree of expression of protein in some protein spots was increased in accordance with decrease of soaking period. Especially, in Hwangkeum-Kong, Danyeop-Kon, and Pecking, the degree of expression of protein was remarkably increased for 4 days after soaking. But, according to germination after soaking, degree of expression of protein in germinated seeds of all cultivars was higher than un-germinated seeds. In results of MALDI-TOF analysis, specific proteins were identified by different soaking period such as Allergen Gly m Bd 28K, P24 oleosin isoform B. Also, in accordance with germination, degree of protein expression of the related protein, Gibberellin was increased in un-germinated seeds of Iksan-Kong. In ungerminated seeds of Sinpaldal-kong, proteins were identified as down-regulated by soaking such as ATP binding and Inhibitor II', proteinase.

In the present study, different expression of protein from Taekwang was revealed by 2-DE, and expressions of protein on each week after flowering was investigated. After analysis of expression of protein, MALDI-TOF was executed to identify expected protein function. Results revealed that there were three patterns of expression of protein during the maturing. The first pattern was that proteins were gradually expressed as up-regulation from 1 week to 6 week. The second pattern was that proteins were expressed gradually from 1 week to 5 week and then it started down-regulation in 6 week. The last pattern was that proteins were gradually as up-regulation from 1 week to 3 week and then down-regulation until 6 week. This phenomenon suggests that young stage has more protein related to correspondence mechanism against disease and growth and then maturing stage has more expression of protein related to storage protein. In MALDI-TOF analysis, p24 oleosin isoform A protein was identified that relates oleosin which is synthetic product in oil body. This protein spot increased gradually until 5 week and then decreased after 5 week. It explained that the protein is active until maturing stage to protect oil in seed and then its activity has gradually degraded. This result may be expected that a protein, related to growth of a seed has increased until maturing and then a seed fills up with a storage protein

Soybean is very useful crop to supply vegetable protein for human. Supply of soybean is increased because it has useful ingredient. Recently, cultivation of soybean in paddy field is increasing due to the increase of rice stockpile in Korea. Hence, in this study, expression of protein was identified regarding different environment for cultivation to investigate the effect of different environment on protein expression. Two-dimensional electrophoresis was performed to investigate the expression of protein using image analysis program to measure degree of protein expression in numerical value. Hannam-kong, Beakcheon-Kong, Hwangkeum-Kong, and Danwon-Kong were used as plant material. 2-DE combined with image analysis revealed that each degree of protein expression of Hannam-Kong and Hwangkeum-Kong in upland field was higher than degree of protein expression in paddy field. However, in case of Beackcheon-Kong, the phenomenon was opposite. In Danwon-kong, the degree of protein expression was not different between up-land field and paddy field. To this end, major protein spots were not different between paddy field and upland field among all cultivars. It could be suggested that protein expression is not severely different by various environment, but different environment affects degree of protein expression.

This study was conducted to improve the postharvest storage techniques of managing and storing seeds, totest qualities and viabilities of the seeds and to examine the germination rate and the protein expression of Achyranthesjaponica Nakai. The seeds collected from different areas of Je-Cheon and Gwang-Ju were stored with different temperaturesand durations. Two plant growth regulators and two seed priming were treated to investigate their effect on the germinationrates and the days required for germination. The weight of one hundred seed collected in Gwang-Ju was heavier than thosein Je-Cheon. Seed length collected in Gwang-Ju was also longer about 5.12㎜ than those in Je-Cheon about 4.90㎜ andseed width was longer in Gwang-Ju than those in Je-Cheon. The rates of seed germination in two different collection areaswere higher about 2.9 to 13.0% in Gwang-Ju compared to those in Je-Cheon. Comparing its rates with the storing tempera-tures and durations, they were not clearly different in between 4℃ and 25℃ and they also were gradually decreased withgetting longer storing durations. The germination rates treated by plant growth regulators were higher with GA3 than thosewith Kinetin. The highest seed germination rate was appeared at 50 ppm of GA3. Comparing its rates with different seedpriming, they were relatively higher with KNO3 than those with PEG6000. In protein expression patterns between before thegerminating and after the germinating of seeds, more and clear bands were appeared in the seed after the germination com-pared to those before the germination of seeds, especially 10~20kDa. These results showing more and clear bands weremore clearly appeared in Gwang-Ju compared to Je-Cheon. Comparing the protein expression with plant growth regulatorsand seed primings, GA3 was better expression than those with Kinetin and KNO3 was better than those with PEG6000. Moreand clear bands were closely related to the germination rates of seeds and more detailed studies would be required.

Serine-arginine-rich nuclear protein LUC7L plays an important role in the regulation of myogenesis in mice. In the present study, we isolated and characterized the Korean rose bitterling Rhodeus uyekii Luc7l cDNA, designated RuLuc7l. The RuLuc7l cDNA is 1,688 bp long and encodes a 364-amino-acid polypeptide containing serine/arginine-rich region at the C-terminus. The deduced RuLuc7l protein has high amino acid identity (71-97%) with those of other species including human. Phylogenetic analysis revealed that RuLUC7L clustered with fish LUC7L proteins. The expression of RuLuc7l mRNA was high in the brain, kidney, and stomach of Korean rose bitterling. Expression of the RuLuc7l mRNA was detected from 1 day post-fertilization (dpf) and moderately increased until 21 dpf during the early development. Further investigations are required to elucidate the functional role of RuLUC7L in myogenesis in R. uyekii.

This study was conducted to investigate the quality of seeds, the germination rates and the days required for germination, to examine the patterns of protein expressions during the germination and to improve the techniques of managing and storing seeds and viability of the seeds of Lithospermum erythrorhizon Sieb. et Zucc. After collecting and harvesting seeds, they were classified to white and brown colors of seed coat through testing their seed size, weight, and quality. The germination rates, the days required for germination, and the protein expressions were examined with different colors of seed coats, storing temperatures and durations by treating the different plant growth regulators and primings. One hundred seed weight of white color was heavier about 1.17 g than those of brown one about 0.81 g. The germination rates in white color of seed coat was higher, 3.05 ~ 5.75%, than those in brown one. Its rates were decreased with getting longer in storage durations. There was no big differences on germination rates between storage temperatures. The plant growth regulator of GA3 and Kinetin was affected to improve the seed germination. GA3 increased the seed germination clearly at 25 ppm level, while kinetin increased it gradually from 25 to 100 ppm levels. In germination by seed primings, PEG6000 made higher germination rate with increasing their levels, whereas KNO3 increased the germination until 100 mM level and then decreased it with 200 mM unlike PEG6000. The protein expressed during the seed germination were appeared more and clearer bands in the seed after germination, especially 20 ~ 30 kDa, compared to those in the seed before germination. These results showing more and clearer bands were positively related to the germination rates which were different by seed colors, storage temperatures and durations, and plant growth regulators and primings.

In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. A cDNA coding for the C-terminus of spider silk protein (AvMaSp) was cloned from the spider Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvMaSp consists of 165 amino acids of are petitive region and 99 amino acids of a C-terminalnon-repetitive region. The peptide motifs found in spider silk proteins, GGX and An, were conserved in the repetitive region of AvDrag. The AvMaSp cDNA was expressed as a 28kDa polypeptide in baculovirus-infected insect cells. To produce transgenic rice plant with high contents of glycine and alanine, the prolamin promoter-driven AvMaSp was introduced into rice plant via Agrobacteriumtumefaciens-mediated gene transformation. Because of seeds pecific prolamin promoter, expression of AvMaSp protein has been achieved inriceseed. The introduction and copy number of the AvMaSp gene in transgenic rice plants were determined by PCR and Southern blot analysis. AvMaSp expression in transgenic rice seeds was examined by Northern blot and Western blot analysis. Immuno fluorescence staining with the AvMaSpantiserum revealed that the recombinant AvMaSp proteins were localized in transgenic rice seeds. Furthermore, the amino acid content analysis showed that transgenic rice seeds were greatly increased in glycine and alanine as compared to controls. The present study is the first to show the expression of spider silk protein in rice seed.