분열효모 S. pombe의 포자형성은 배지상의 질소원 고갈에 의해 유도되어지며 감수분열로부터 포자형성에 도달하는 과정에는 다수의 특이적인 유전자들이 관여하고 있다. 본 실험에서는 S. pombe genomic library 형질 전환법으로 spo5 유전자를 상보하는 clon을 screening 한 후, spo 5 유전자를 단리하였다 ^8). 전포자막 구축에 필수적인 spo 5 유전자를 보유하는 약 5kb의 DNA 단편을 대장균, 효모 shuttle vector pTB248의 Hind Ⅲ 부위에 subclonning 하였다. 그리고 이 DNA 단편으로부터 제한효소 지도를 작성하여 (Fig-2), spo 5 변이체의 상보능력을 조사하였다. (Fig 3). 결과에서 서술한 바와 같이 상보능력은 동일하였으며, 이러한 상보성 실험 결과로부터 삽입된 단편상의 유전자 발현은 벡터의 promoter로 부터 전사가 일어나는 것이 아니라, 삽입 단편상의 효모 고유의 promoter 에 의해서 전사가 일어나는 것으로 확인되었다. 따라서 clone화 한 DNA 단편 배열상에는 변역영역뿐만 아니라 promoter 영역이 포함된 것으로 판단되었다. 결실변이 도입 해석으로부터, spo 5 유전자는 Sma I부터 Hind Ⅲ의 3kb 영역에 존재하였고 (Fig 2), Northem 분석에 의해서 spo5-mRNA는 Sma Ⅰ부터 Hind Ⅲ의 3kb 영역에서 약 2.5kb 크기로 검출되었다. 이 단편의 유전해석으로부터 약 2.5kb 크기로 검출되었다. 이 단편의 유전해석으로부터 약 2.5kb의 전사산물은 최대 800개의 아미노산 잔기를 code하는 단백질로 판단되었다. (Fig 4). 그리고 , Northem 분석법에 의해서 spo 5 유전자의 전사를 조사한 결과, 서술한 바와 같이, 이 유전자는 질소기아 조건 하에서 만 유전자가 발현되는 것을 확인하였다. (Fig 4 -2.5kb 단편). 그 결과, 최소배지의 hetro matting-type 균주 (CD16-1) (Table 1)로 부터 조제한 mRNA와 probe-DNA가 hybridize하는 전사산물은 spo 5 유전자 coding 영역에서 3.2kb와 2.5kb의 mRNA가 검출되었다 (Fig 4). 그리고 homo matting -type (CD16-3) (Table I) 균주를 질소원이 함유되지 않은 포자형성 배지에서 배양한 후, 동일한 방법으로 mRNA를 조제하여 Northem hybridization으로 조사하였다. 그 결과, 이들 세포에서는 3.2kb에서 만 전사산물이 검출되었으며, 2.5kb의 mRNA는 검출되지 않았다. (Fig 4). 이상의 결과로부터 spo 5 유전자를 coding 하는 전사산물인 2.5kb의 mRNA는 질소원이 고갈된 상태하에서, 접합형 유전자좌의 hetero 접합성을 요구하는 것으로 입증되었고, spo 5 유전자의 전사발현은 질소원이 결핍과 집합형 유전자좌의 구성에 따른 환경요인과 유전적 요인에 의해서 제어되어지고 있다는 것을 입증하였다.

흰쥐의 체중은 실험기간 동안 증가하였으며, 식이별로는 대조군이 가장 높은 체중 증가를 보였다. 콜레스테롤의 첨가는 흰쥐의 정상적인 성장을 억제하였으며, 콜레스테롤 첨가군에서는 콜레스터롤 대조군의 체중증가량이 유의하게 가장 낮았다. Rutin과 메밀채소를 식이에 첨가하였을 때 흰쥐의 콜레스테롤에 의한 성장억제 작용을 효과적으로 감소시키는 것으로 나타났으며, 첨가량의 증가에 따라 헐청에 총콜레스테롤 함량과 과산화지질 함량은 낮아진 반면에 HDL-cholesterol 함량은 높아지는 경향을 보였다. 고콜레스테롤 식이를 급여시킨 흰쥐의 장기에서는 칼슘침착이 나타나는 병리조직학적 변화를 보였으며, 메밀채소 섭취에 의하여 칼슘침착의 정도는 완화되었다.

청둥오리의 압란유의 기능성 특성을 알아보기 위하여 흰쥐를 모델로 하여 streptozotocin(STZ)을 투여하여 당뇨병을 유발시킨 다음, 혈액 중의 당 농도, 지질의 변화 및 동맥경화 지표와의 상관관계를 검토하였고, 또한 Sarcoma-180을 쥐에 이식하여 이에 대한 항암 효과를 실험하였다. STZ로 당뇨병을 유발시킨 흰쥐를 청둥오리의 압란유를 15일간 투여하였던 바 혈중의 당 농도는 정상상태로 유지되었고 phospholipid 및 triglyceride의 함량은 STZ를 처리한 압란유의 경우에는 증가하였으나, STZ 처리군에 압란유를 투여하였을 때 감소되었다. 혈중 total cholesterol, LDL+VLDL의 cholesterol 및 동맥경화의 지표는 STZ 처리군에서는 증가하였으나, 시료를 150㎎/㎏ 투여시에는 감소하였다. 한편 Sarcoma-180에 대한 성장억제율은 압란유를 150㎎/㎏ 투여시 63.89%로 나타났고 수명연장 실험에서는 압란유의 경우 15.4%로 나타났다.

Bacillus cereus B-amylase was purified by Sephadex G-100 gel filtration, CM Sephadex C-50 ion exchange chromatography and CM Sephadex C-50 ion exchange rechromatography. The purified enzyme showed 871unit/㎎ of specific activity. The purified enzyme was identified as homogenious by disc PAGE, SDS-PAGE and analysis of reaction product. The purified enzyme showed optimum pH 7.0. optimum temperature 50℃, and was stable at 0∼50℃ and at pH range of 6∼10.

This research aims to find the information concerning nutritional status and food intake in Korean diabetic men. Thus, the survey was made on 202 NIDDM patients over 35 age at University Hospital. The data were analysed using F-test and mutiple comparison in SAS package program. Main results were as follows: Incidence of diabetes is high in their forties and fifties. Most of them are salary men, administrators, and proffesionals in middle class, who reside in metropolitan area including Seoul. They used to take light exercise and were founded NIDDM by subjective symptoms. 36.6% of them have other diseases simultaneously such as hypertension, cancer, etc. Food intake pattern prior to incidence was that intake frequency of grains, sugars, protein foods, and liking foods was higher than of proccesed foods. Amounts of calorie intake were shown to be lower than normal persons, so that it seemed to be controlled by doctors. In addition, the calorie ratio derived from fat was lower than RDA. Thus protein and carbohydrate ratio was relatively higher than RDA. Total mean fatty acid intake was 32.15g and it is no significance among weight groups in various fatty acid intakes. With respect to the physical standards of the diabetic men, weight, PIBW, BMI, BMM, TST, arm circumference, and waist/hip ratio were higher than in normal. Therefore it seemed that these factors would be related to the diabetics. From now on, it is reqiured to research the correlations of fatty acid intakes on the diabetes and their influence to serum lipid profiles.

To examin ripening in peach types, cell wall contents and Polygalacturonase activity were compared in Changbang, Daegubo and Yumyung peaches. Among peach types, the hardness of Daegubo was the lowest. Yumyung peach had the highest content of alcohol-insoluble substances and Changbang peach of cell wall. The contents of total and insoluble pectic substances were little different between Changbang and Yumyung peach, while the lowest in Daegubo. Daegubo peach had the highest activity of polygalacturonase, Changbang and Yumyung peach in succession.

Effects of metal chelating agents and metal ions on the volatile substance of cooked oil with chicken and pork mixture meat were examined by chemical analysis and sensory test. The addition of Na-tripolyphosphate(Na-TPP) to chicken and pork mixture meat increased the amount of H_2S among volatiles evolved during cooking but decreased that of volatile carbonyl compounds(VCC). This treatment enhanced meat flavor in cooked oil with chicken and pork mixture. It was recognized that the increase in H_2S evolution was caused by the rise of pH value. On the contrary cupric ion produced a negative effect on the production of chicken and pork mixture meat flavor and this addition increased VCC and TBA value. Other metal chelating agents such as citric acid, phytic acid and EDTA, provided the same results as Na-TPP. It was supposed that these phenomena were attributable to the chelating action to metal prooxidant in mixture meat. It could be concluded that a proper evolution of H_2S and protection against lipid oxidation during cooking were important to produce an excellent chichen and pork mixture meat flavor.

본 논문의 내용은 혈간콘크리트 보-기둥 접합부가 지진 하중을 받을때의 거동에 대하여 관찰한 것 이다. 똑같이 제작된 두개의 시험체에 정적 반복하중과 동적반복하중을 가하여 하중-처집 곡선이나 파괴 성상동에 판하여 차이점을 기록하였다. 동적하중을 받는 시험체의 거동은 내진설계 규준에서일 반적으로 쓰여지는 정적 하중 하에서의 시험체의 거동과는 판이한 양상올 보여주었다. 시험체가 동적 하증을 받을때에는 정적하중올 받을때 보다 @ 극 한하중이 20-25 % 증 가하고 @ 높은 취성을 보이며 @ 균열이 집중되고， @ 휩파괴 보다는 전단파괴현상을 나타내었다.

본 연구는 절임배추의 유통 중 단기 유통시스템에 따른 이화학적 및 미생물학적 품질 변화에 대하여 알아보고, 그에 따른 품질지표를 평가하고자 실시하였다. 여름철에 제조한 절임배추는 탈수 후 바로 저밀도 폴리에틸렌 필름에 포장하거나 2% 염수와 함께 포장하여 실온유통시스템(conventional system)과 저온유통시스템(cold-chain system)을 갖춘 1톤 트럭에 각각 구분하여 적재하였으며 약 6시간동안 유통하였다. 이화학적 및 미생물학적 변화로 품온, 염도, 산도, pH, 수분감, 일반세균, 젖산균, 대장균, 대장균군, 효모 및 곰팡이에 대해 조사하였다. 실온유통시스템의 경우 절임배추의 품온은 필름 내 염수포장 절임배추에서 19.57℃, 필름포장 절임배추에서 19.43℃, 저온유통시스템의 경우 필름 내 염수포장 절임배추에서 10.73℃, 필름포장 절임배추에서 12.90℃까지 증가하였다. 염도변화는 저온유통시스템의 경우 변화가 없었으나, 실온유통시스템의 경우 필름 내 염수포장 절임배추와 필름포장 절임배추에서 품온과 각각의 염도변화가 정의 상관성을 보이며 초기에 비해 1.2배 혹은 1.7배 더 높았다. 산도는 저온유통시스템에서 두 가지 포장조건 모두 약간 증가하였다. 실온유통시스템에서 필름 내 염수포장 절임배추의 총 호기성균과 젖산균수가 각각 7.62 log CFU/g 및 6.77 log CFU/g까지 증가하는 동안 필름포장 절임배추는 각각 5.62-5.85 log CFU/g과 4.33-4.83 log CFU/g의 범위를 나타냈다. 그러나 저온유통시스템의 경우 유통시간 증가에 따른 유의미한 미생물학적 변화를 보이지 않았다. 따라서 절임배추 유통 시 저온유통시스템을 이용한 경우 이화학적 품질 유지 및 미생물 제어에 효과적이며, 실온유통시스템을 이용한 경우 품온 상승에 따른 염도 및 미생물 변화에 유의해야 한다. 또한 염도, 총호기성균 및 젖산균은 실온유통시스템 내 품질지표로 이용가능하며, 산도는 저온유통시스템 내 품질지표로 이용가능하다.

High yield is the most important trait in various agricultural characteristics. Many approaches to improve yield have been tried in conventional agricultural practice and recently biotechnological tools employed for same goal. Genetic transformation of key genes to increase yield is one way to overcome current limitation in the field. We are producing transgenic soybean plants through high efficient transformation method by introducing all gene member with AT-hook binding domain, hoping to obtain manageable delay of senescence. Many transgenic soybean plants are growing in greenhouse and GMO field, and will be evaluated their senescence and any association with yield increase.

새로운 베지클인 glyceryl citrate/ lactate/ linoleate/ oleate를 이용한 수중유형 형태의 아스타잔틴 나노에멀젼에 대해 항산화 효과, 세포 생존력, 단백질과 관련한 효소의 영향, 피부 침투도 그리고 피부에 대한 보습 및 탄력 등의 약용화장품적인 측면에서의 전반적 연구를 실시하였다. 항산화력 및 세포 생존력에 대해선 각각 DPPH법과 MMT assay를 이용하여 측정하였다. 아스타잔틴 나노에멀젼에 대한 또 다른 성질은 2D-Page를 이용한 단백질 분석 및 컨포칼, in-vivo 테스트를 통해 측정하였다. 본 연구를 통해, 아스타잔틴을 포함하는 나노에멀젼은 MMP발현에 관련한 단백질 억제 및 세포외 기질의 분해를 막고 라디칼의 소거에 매우 우수한 결과를 보였다. 종전의 레시친을 이용한 나노에멀젼 보다는 새로운 베지클을 이용한 아스타잔틴 나노에멀젼의 피부 침투가 매우 효과적임을 CLSM을 통해 측정하였다. 또한 28일 동안의 한국 성인 여성 11명을 통한 보습 및 탄력 인비보 테스트에서 우수한 효과를 확인할 수 있었다.

AtRabG3b and CaMsrB2 genes incorporated into pPZP vetor were transformed to Korean soybean cultivar Kwangan using highly efficient transformation system. AtRabG3b gene plays a positive role in xylem development in Arabidopsis and 64 transgenic plants were produced. CaMsrB2 gene is known to confer drought tolerance in rice and 63 transgenic plants were produced. As a result of PPT leaf painting assay, about 20% of transformation efficiency was observed from 2 times of inoculation. These transgenic plants were confirmed for gene introduction using PCR. Currently, the copy number and the gene expression is investigating using qRT-PCR and RT-PCR. Moreover, 62 lines and 53 lines of T1 seeds from AtRabG3b and CaMsrB2, respectively, were sown in GMO field.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Totally, 26 collections, 17 from Korea and 9 from China, were investigated for their sequences of 5S rDNA, especially the non-transcribed spacers (NTSs). Sequences of 5S rDNA were isolated by PCR using the primers, 5s-rRNA1 and 5s-rRNA2. Genomic DNA PCR produced single amplification of 300, 330, or 350 base pair fragments. Sequence analysis revealed that all inserts contained the part of 5S rDNA gene sequence and the full length of the NTS region. Three different sizes of the fragments were confirmed due to different size of NTS and their length were 300bp, 330bp and 350bp, respectively. Among 17 Korean foxtail millets tested, 14 collections showed single 300bp amplification. Longest fragment amplification, 350bp, was obtained only from the foxtail millet from China origin, even though 2 of them include 300bp fragment. CLUSTALW multiple alignments of 26 foxtail millets clearly revealed 4 areas with certain degree of sequence heterogeneity (region I, II, III, IV). Among 4 boxed areas, foxtail millet genotypes from China have distinct insertion especially in region III. Five of them have extra insertion of sequence and their additional sequences were either 45 or 48 base pair. Three Korean foxtail millets have 32 bp insertion. Other 8 Korean collections have short insert sequences (6 to 8 bp), 3 with 8 bp and 5 with 6 bp. In addition to insert, deletion sequences were also confirmed as major deletion was observed in region II of Chinese collection. The size of deletion was 7 bp long. According to phylogenic tree constructed using MEGA4 program, clear grouping was not revealed. To obtain more convincing results various collections from many countries should be obtained and analyzed to distinguish different germplasm from different origin.

Twenty two common millet (Panicum miliaceum L.) varieties collected from Korea, China and Russia were investigated for their phylogenetic relationship using 5S ribosomal DNA sequences with a hope to provide the basic information on their exact origin. Sequences of 5S rDNA were isolated by PCR. The primers, 5s-rRNA1 and 5s-rRNA2, were designed to isolate the complete NTS. Genomic DNA amplification produced two fragments with different length, 900 bp and 400 bp fragments, confirming the presence of two types of 5S rDNA repeats that differed from each other in the length of the NTS region. Amplified DNAs of 400 bp fragment were subcloned and used for further investigation. The obtained NTS sequences ranged from 200 to 300 bp and homology of sequences among plant materials was much higher than long repeat. CLUSTALW multiple aligment of 5S rDNA sequences from 22 different common millets revealed the clear difference by their origin. And critically different areas with insert or deletion were also confirmed. Those sequence difference seems to be used for discrimination of cultivars from different origin and use as molecular markers for origin identification. In phylogenic tree construction, the clear classification was shown where the genotypes from China and Russia is positioned together and stay away from domestic genotypes.

Korean soybean variety Kwangan was transformed with coat protein (CP), helper component-proteinase (HC-Pro), and ABRE binding factor 3 (ABF3) genes using highly efficient soybean transformation system. Among these genes, CP and HC-Pro were transformed using RNAi technology. Transgenic plants with CP were confirmed for gene introduction and their expression using PCR, real-time PCR, RT-PCR, Southern blot, and Northern blot. To investigate the response of viral infection with CP, T1 plants were inoculated with SMV-infected leaves and confirmed the existence of mosaic symptom in both leaves and seeds. Two transgenic lines with CP were highly resistant to SMV with clear leaves and seeds while SMV-susceptible lines showed mosaic symptom with seed mottling. The transcript levels of T1 plants with CP were also determined by northern blot, suggesting that SMV-resistant T1 plants did not show viral RNA expression whereas SMV-susceptible T1 plants showed viral RNA expression. Currently, the response of viral infection with HC-Pro is investigating to produce SMV-resistant soybean transgenic plants, and the physiological experiment with ABF3 is also carrying out to produce drought-tolerant soybean transgenic plants.

Wild rice might have previously unidentified genes important for disease resistance and stress tolerance in response to biotic and abiotic stresses. A set of subtractive library was constructed both from leaves of wild rice plants, Oryza grandiglumis (CCDD, 2n=48), treated with fungal elicitor and from wounded leaves. A partial fragment that was homologous to PR10 genes from other plant species was identified via suppression subtractive hybridization and cDNA macroarray. The obtained full-length cDNA sequence (OgPR10) contains an open reading frame of 480 bp nucleotide, encoding 160 amino acids with a predicted molecular mass of 16.944 kDa and an isoelectric point (pI) of 4.91. The multiple alignment analyses showed the higher sequence homology of OgPR10 with PR10 genes identified in rice plants at amino acid level. The OgPR10 mRNA was not expressed by treatment with wounding, jasmonic acid, and salicylic acid, but markedly expressed in leaves treated with protein phosphatase inhibitors cantharidin and endothall, and yeast extract. In addition, the expression of OgPR10 mRNA was induced within 72 h after treatment with probenazole, one of well-known chemical elicitors, and reached the highest level at 144 h. Heterologous expression of OgPR10 caused growth inhibition and seedling lethality in E. coli and Arabidopsis, respectively. Chemically induced OgPR10 expression with glucocorticoid-mediated transcriptional induction system further reconfirmed its lethality on Arabidopsis seedling. In addition, OgPR10-expressing rice plants, Oryzae sativar were resistant against the infection of rice blast fungus, Magnaporthe grisea. These results indicate that OgPR10 is involved in probenazole- and microbe associated molecular patterns-mediated disease resistance responses in plants and is a potential gene for developing disease resistance crop plants.