• Title/Summary/Keyword: Expression rate

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Ethanol extract of Aster glehni exhibits anti-inflammatory and anti-oxidant effects in RAW 264.7 cells and Caenorhabditis elegans (섬쑥부쟁이 에탄올 추출물이 대식세포와 예쁜꼬마선충에서의 항염증 및 항산화 효과 )

  • Mi-Kyung Seo;Han-Na Chu;Da-Bin Lee;Haeng-Ran Kim;In-Seon Hwang;Yong-Jin Jeong;Sung-Ran Yoon;Seok-Seong Kang;Kyeong-A Jang;Min-Sook Kang
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1095-1106
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    • 2023
  • This study investigated the anti-oxidative and anti-inflammatory effects of Aster glehni (AG) extract in RAW 264.7 cells and Caenorhabditis elegans. The total polyphenol and flavonoid contents were higher in the ethanol extracts than in the hot water extracts. As a result of measuring the moisture contents (%) and extraction yields (%) of AG and drying A. glehni for processing (DAG), 70% ethanol, which has the highest percentage of extraction yield, was selected as the final solvent. DPPH radical scavenging activity showed higher antioxidant activity of ethanol extracts of DAG than AG. The cytotoxicity assay of the AG or DAG ethanol extracts was treated at different concentrations (25, 50, and 100 ㎍/mL), and cell viability rates were higher than 80% at all concentrations. The LPS-stimulated nitric oxide (NO) production in RAW 264.7 was significantly reduced at all concentrations of AG and DAG groups. As a result of measuring the gene expression of iNOS, which induces NO production, the AG or DAG group decreased by 33% and 32%, compared with the phosphate buffer saline (PBS) group. Under inflammatory stress conditions, the survival rate of C. elegans treated with AG or DAG ethanol extract with LPS showed concentration-dependent improvement in survival rate compared with the PBS group. Considering these results, AG could potentially be developed as an antioxidant and anti-inflammatory functional food material.

Promoter Polymorphism of RRM1 Gene in Korean Lung Cancer Population (한국인 폐암 환자에서 RRM1 유전자 Promoter의 다형성)

  • Ko, Kyung Haeng;Kim, Eun Joung;Oh, In Jae;Kim, Soo Ock;Son, Jun Gwang;Jung, Jong Pil;Cho, Gye Jung;Ju, Jin Young;Kim, Kyu Sik;Kim, Yu Il;Lim, Sung Chul;Kim, Young Chul;Bepler, Gerold
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.3
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    • pp.248-255
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    • 2006
  • Background: LOH11A is a region with frequent allele loss (>75%) in lung cancer that is located on the centromeric part of chromosome 11p15.5. Clinical and cell biological studies suggest that this region contains a gene associated with metastatic tumor spread. RRM1 encoding the M1 subunit of ribonucleotide reductase, which is an enzyme that catalyses the rate-limiting step in deoxyribonucleotide synthesis, is located in the LOH11A region. Methods: Polymorphisms were found at nucleotide position (-)37 (C/A) and (-)524 (C/T) from the beginning of exon 1 of the RRM1 gene that might regulate the expression of RRM1. We studied the polymorphisms in 127 Korean individuals (66 lung cancer and 61 normal controls) and compared with those of 140 American patients with lung cancer. Results: CC, AC and AA were found at the (-)37 position in 64(50.4%), 55(43.3%), and 8(6.3%) out of 127 Korean individuals (66 cancer, 61 non-cancer patients), respectively. There was a similar frequency of allele A at (-)37 in the American(27.9%) and Korean population(28.0%). CC, CT and TT was found at the (-)524 position in 24(18.9%), 44(34.6%), and 59(46.5%) out of the 127 Korean individuals, respectively. There was a similar frequency of allele C at (-)524 in the American(34.6%) and Korean population(36.2%). There was no difference in the frequency of the (-)37 and (-)524 genotypes between the cancer and non-cancer group. However there was a significant correlation of the genotypes between (-)37 and (-)524 (p<0.001), which suggests the possible coordination of these polymorphisms in the regulation of the promoter activity of the RRM1 gene. Conclusion: RRM1 promoter polymorphisms were not found to be significant risk factors for lung cancer. However, a further study of the promoter activity and expression of the RRM1 gene according to the pattern of the polymorphism will be needed.

Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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The Validation Study of the Questionnaire for Sasang Constitution Classification (the 2nd edition revised in 1995) - In the field of profile analysis (사상체질분류검사지(四象體質分類檢査紙)(QSCC)II에 대(對)한 타당화(妥當化) 연구(硏究) -각(各) 체질집단(體質集團)의 군집별(群集別) Profile 분석(分析)을 중심(中心)으로-)

  • Lee, Jung-Chan;Go, Byeong-Hui;Song, Il-Byeong
    • Journal of Sasang Constitutional Medicine
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    • v.8 no.1
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    • pp.247-294
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    • 1996
  • By means of the statistical data which has been collected with newly revised QSCC made use of the outpatient group examined at Kyung-Hee Medical Center and an open ordinary person group, the author proceeded statistical analysis for the validation study of the revised questionnaire itself. First, check the accurate discrimination rate by performing discriminant analysis on the statistical data of the patient group. And next, sought T-score by applying the norms gained in process of standadization of the open ordinary person group to the Sasang scale score of the outpatient group and investigated the distinctive feature between the subpopulations which was devided in the process of multivarite cluster analysis. The result was summarized as follows ; 1. The validity of the questionnaire was established through the fact that the accurate discrimination rate the ratio between predicted group and actual group was figured out 70.08%. 2. At the profile analysis the response to the relevant scale showed notable upward tendency in each constitutional group and therefore it seems to be pertinent in the field of constitutional discrimination. 3. In the observation of the power of expression through the profile analysis of each constitutional group the Soyang group demonstrated the most remarkable outcome, the Soeum group was the most inferior and the Taieum group revealed a sort of dual property. 4. What is called the group of seceder out of three subpopulation of each constitutional group distinguished definitely from the contrasted groups at the point of the distinctive profile feature and the content is like following description. (1) The seceder group of Soyang-in showed considerably passive disposition differently from general character of ordinary Soyang group and an appearance attracting the attention is that they demonstrated comparatively higher response at Soeum scale (2) The seceder group of Taieum-in gained low scores in general that informed the passive disposition of the group and the other way of the general property of Taieum group which showed accompanied ascension in Taiyang-Taieum scales they demonstrated sharply declined score at Taiyang scale (3) The seceder group of Soeum-in demonstrated distinctive property similar to the profile feature of Soyang group and it notifies that the passive property of Soeum group was diluted for the most part. According to the above result, the validity of newly revised questionnaire has been proven successfully and the property of seceder groups could be noticed to some degree through the profile analysis on the course of this study. The result of this study is expected to use as a research materials to produce next edition of the questionnaire and it is regarded that further inquisition about the difference between the seceder group and the contrasted group is required for the promotion of the questionnaire as it refered several times in the contents of the main discourse.

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Effect of retinoic acid on the bystander effect in gene therapy using the Herpes Simplex Virus thymidine kinase (Herpes Simplex Virus thymidine kinase gene을 이용한 유전자 치료에서 retinoic acid가 bystander effect에 미치는 영향)

  • Park, Jae Yong;Kim, Chang Ho;Jung, Tae Hoon;Albelda, Steven M.
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.1
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    • pp.162-174
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    • 1997
  • Background : Metabolic cooperation via gap junctional intercellular communication (GJIC) is an important mechanism of the bystander effect in gene therapy using the Herpes Simplex Virus thymidine kinase/ganciclovir (HSVtk) "prodrug" system. Since retinoids have been reported to increase GJIC by induction of connexin 43 expression, we hyporthesized that treatment of tumor cells with retinoic acid could augment the bystander effect of the HSVtk/GCV system and result in improved tumor cell killing by enhancing GJIC. Methods : We transferred HSVtk gene to SKHep-J cell line that does not express connexin43, and also transferred the gene to human and murine mesothelioma cell lines that express connexin43. We verified that retinoic acid enhanced GJIC utilizing a functional double-dye transfer study and evaluated the effects of retinoic acid on the growth rate of tumor cells. We then tested the effects of retinoic acid on bystander-mediated cell killing. Results : Addition of all-trans retinoic acid (RA) increased GJIC in cell lines expressing connexin 43 and was asspciated with more efficient in vitro bystander killing in cells transduced with HSVtk via adenoviral and retroviral vectors. In contrast, there was no increase in the efficiency of the bystander effect after exposure to RA in a cell line which had no delectable connexin 43. Conclusion : These results provide evidence that retinoids can augment the efficiency of cell killing with the HSVtk/GCV system by enhancing bystander effect and may thus be a promising new approach to improve responses in gene therapy utilizing the HSVtk system to treat tumors.

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Evaluation on Functional Assessment for Fish Habitat of Underground type Eco-Artificial Fish Reef using the Index of Biological Integrity (IBI) and Qualitative Habitat Evaluation Index (QHEI) (생물보전지수(IBI) 및 서식지 평가지수(QHEI)를 활용한 지하 매립형 방틀둠벙의 어류 서식처 기능 평가)

  • Ahn, Chang Hyuk;Joo, Jin Chul;Kwon, Jae Hyeong;Song, Ho Myeon
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.31 no.6B
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    • pp.565-575
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    • 2011
  • The purpose of this study was to quantitatively evaluate the expression of both multi-metric qualitative habitat evaluation index (QHEI) and biological integrity index (IBI) for artificial structures eco-artificial fish reef (EAFR) for fishes asylum and habitat. Especially, both experimental evaluation and biological verification were performed in Water and Environmental Center's outdoor test-bed of Korea Institute of Construction Technology located in Andong-city, Gyeongsangbuk-do. The experimental conditions reflecting the situation of domestic river include the flow rate (e.g., $0.0{\sim}1.5m\;s^{-1}$), the width (e.g., 1.0~3.0 m), the depth (e.g., 0.05~0.70 m), and variable bed materials. Both QHEI and IBI were monitored for 8 months from May to December 2010. Whereas QHEI values were highest at experimental points of the E~F with an average of 83.1, those were lowest at B~C with an average of 78.1. However, QHEI values inside EAFR were more than 98.9, regardelss of space and time, and indicated more than the highest good of the state (Good) in the habitat. Overally, IBI values showed similar trend with QHEI, but were 44.2 in the winter dry season, compared to 32.8 of QHEI values. IBI values Also, IBI values inside EAFR were greater than those at the experimental channel by 5.7 to 11.4% and 18.7 to 34.8% in flow and stagnant conditions, respectively, indicating that EAFR can secure asylum and habitat for fish during the dry season. For comprehensive aquatic ecosystem assessment, the experimental channel showed generally fair conditions (Fair~Good), whereas EAFR showed good conditions (Good), suggesting that EAFR can be applied to aquatic ecosystem restoration and improvement.

Enhancement of β-1,3-Glucanase Activity by Sequential δ-Sequence Mediated Integration in Saccharomyces cerevisiae (출아효모에서 연속적 δ-sequence 삽입유도에 의한 β-1,3-glucanase 활성 증가)

  • Kim, Min-Jung;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1046-1054
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    • 2014
  • Beta-1,3-glucanase is widely used in various biotechnological and industrial processes, with over-production required to enable versatile utilization. We examined the overexpression of ${\beta}$-1,3-glucanase (EXGA) from Aspergillus oryzae using ${\delta}$-sequence-mediated integration. We constructed $pRS{\delta}$-exgA and $pRS{\delta}K$-exgA plasmids for integration of the EXGA gene into various chromosomes of Saccharomyces cerevisiae. These plasmids contain the ADH1 promoter for constitutive expression, a signal sequence (exoinulinase signal sequence [INU1 s.s]) for secretory production, and a ${\delta}$-sequence for integration of ${\beta}$-1,3-glucanase. The $pRS{\delta}$-exgA plasmid was transformed into the S. cerevisiae $BY4742{\Delta}exg1$ strain, and ${\beta}$-1.3-glucanase was stably overexpressed and secreted. Another plasmid, $pRS{\delta}K$-exgA, was introduced into the S. cerevisiae $BY4742{\Delta}exg1$ (YKY082) strain, and overexpression of ${\beta}$-1,3-glucanase was examined by inducible integration under geneticin selection. The activity of ${\beta}$-1,3-glucanase increased in accordance with a rise in the geneticin concentration, with 0.8 mg/ml of geneticin suitable for overexpression of ${\beta}$-1,3-glucanase. Subsequently, $pRS{\delta}K$-exgA was repeatedly transformed for sequential ${\delta}$-integration. The activity of ${\beta}$-1,3-glucanase reached about 0.063 unit/ml/$OD_{600}$, 0.095 unit/ml/$OD_{600}$, 0.131 unit/ml/$OD_{600}$ and 0.165 unit/ml/$OD_{600}$ by the first, second, third, and fourth round of integration, respectively. According to the increase in the activity of ${\beta}$-1,3-glucanase by sequential ${\delta}$-integration, the copy number (integration rate) of the EXGA gene also increased in various chromosomes. These results suggest that recombinant ${\beta}$-1,3-glucanase activity can be sequentially increased by repeated ${\delta}$-sequence integration.

Transformation of Bottle Gourd Rootstock (Lagenaria siceraria Standl.) using GFP gene (GFP유전자를 이용한 대목용 박 형질전환)

  • Lim, Mi-Young;Park, Sang-Mi;Kwon, Jung-Hee;Han, Sang-Lyul;Shin, Yoon-Sup;Han, Jeung-Sul;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.33-37
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    • 2006
  • Bottle gourd (Lagenaria siceraria Standl.) has been used as a rootstock for the watermelon cultivation because of better growth ability at low temperature and avoidance from contamination of the soil disease. Since the genetic source for the elite rootstock is limited in nature, the genetic engineering method is inevitable to develop new lines especially to obtain the functionally important or multi-disease resistant bottle gourd. Recently, our lab has set up a successful system to transform the bottle gourd. in order to monitor the transformation process, GFP gene is used. Cotyledons of the inbred line 9005, 9006 and G5 were used to induce the shoot under the selection media with MS + 30 g/L sucrose + 3.0 mg/L BAP + 100 mg/L kanamycin + 500 mg/L cefotaxime + 0.5 mg/L $AgNO_3$, pH 5.8. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. The shoot was incubated in the rooting media with 1/2 MS + 30 g/L sucrose + 0.1 mg/L IAA + 50 mg/L kanamycin + 500 mg/L cefotaxime, pH 5.8 and moved to pot for acclimation. Although the shoot development rate was depended on the genotype, the G5 was the best line to be transformed. Monitoring GFP expression from the young shoot under microscope could make the selection much easier to distinguish the transformed shoot from the non-transformed shoots.

Production of Medium-chain Fatty Acids in Brassica napus by Biotechnology (유채에서의 중쇄지방산 생산)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Kim, Jong-Bum
    • Journal of Applied Biological Chemistry
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    • v.53 no.2
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    • pp.65-70
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    • 2010
  • Medium-chain fatty acids (MCFA) are composed of 8-12 carbon atoms, and are found in coconut, cuphea, and palm kernel oil. MCFA were introduced into clinical nutrition in the 1950s for dietary treatment of malabsorption syndromes because of their rapid absorption and solubility. Recently, MCFA have been applied to Gastrointestinal Permeation Enhancement Technology (GIPET), which is one of the most important parts in drug delivery system in therapeutics. Therefore, to accumulate the MCFA in seed oil of rapeseed, much effort has been conducted by classical or molecular breeding. Laurate can be successfully accumulated up to 60 mol% in the seed oil of rapeseed by the expression of bay thioesterase (Uc FatB1) alone or crossed with a line over-expressing the coconut lysophosphatidic acid acyltransferase (LPAAT) under the control of a napin seed-storage protein promoter. Also, caprylate and caprate were obtained 7 mol% and 29 mol%, respectively, from plants over-expressing of the medium-chain specific thioesterase (Ch FatB2) alone or together with the chain-length-specific condensing enzyme (Ch KASIV). Despite the success of some research in utilizing parallel classical and molecular breeding to produce MCFA, commercially available seed oils have for the most part, not been realized. Recent research in the field of developing MCFA-enriched transgenic plants has established that there is no single rate-limiting step in the production of the target fatty acids. The purpose of this article is to review some of the recent progress in understanding the mechanism and regulation of MCFA production in seed oil of rapeseed.

Evaluation of Scab Resistance and Effect of Photosynthetic Rates on Fruit Characteristics among Elite Pear Seedlings (배 우량계통의 검은별무늬병 저항성 평가 및 광합성률이 과실 품질에 미치는 영향 구명)

  • Won, Kyung-Ho;Kang, Sam-Seok;Kim, Yoon-Kyeong;Sherzod, Rajametov;Lim, Kyeong-Ho;Lee, Han-Chan
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.117-122
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    • 2013
  • BACKGROUND: The scab, which is caused by Venturia nashicola, gives serious damages to pear trees. 'Niitaka' accounts for 82% of areas in pear cultivation. However 'Niitaka' is a scab susceptible cultivar. So, most of Korean farmers who growing pear trees have suffered by economic losses with the scab. In this research, we evaluated the scab resistance among elite pear seedlings to clarify genetics about the scab resistance. And we analyzed photosynthetic features with these seedlings to develop suitable cultivar which is advantageous for producing quality fruits during the growth and development of plants. METHODS AND RESULTS: We measured the rates of scab incidence among seedlings in a field experiment condition and an in-vitro test. An in-vitro test has been done with field experiment-based results. We made plant materials by grafting branches of each seedlings with 'Kongbae' rootstocks. And they had been grown for one month. Then, scab conidia suspension is sprayed to seedlings and sustained for 40 days under the controlled environment. As the results, 6 seedlings displayed lower incidence rates than other seedlings and 'Niitaka'. We also measured instant photosynthetic rates of each seedlings to determine the correlation between photosynthetic rates and fruit characteristics. However, it seemed that there is no correlation between them. CONCLUSION(S): Among the seedlings, 6 seedlings displayed the higher resistance to scab than other seedlings and 'Niitaka'. This characteristics is considered to be come from the gene expression of European pear. And we found that photosynthetic rate in trees rarely does not influence the fruit characteristics. It is considered to be affected by cultivar's own characteristics.