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Purification and Characterization of Protease Produced by Aspergillus wentti Isolated from Korean Traditional Meju (한국 전통 메주 유래의 Aspergillus wentti가 생성하는 Protease 의 정제 및 특성)

  • Lim, Seong-Il
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.161-167
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    • 2000
  • The protease produced by a newly isolated Aspergillus wentti from Korean traditional Meju was purified and characterized. The optimal medium composition and culture conditions for maximum protease production were ; bran :1% glucose solution =1 : 1, pH 9.0, $30^{\circ}C$, and 4 days of fermentation. Protease was purified by QAE-Sephadex, SP-Sephadex ion exchange chromatography and Sephadex G-100 chromatography. The specific activity and the purification fold of the purified enzyme were 213 unit/mg protein and 27.3, respectively. The molecular weight of purified protease was found to be 32 kDa by SDS-PAGE. Km and Vmax value's for hammastein milk casein were $3.049{\times}10^{-4}\;M\;and\;151.1\;{\mu}g/min$, respectively. Kinetic parameters showed that the enzyme has higher affinity to casein than isolated soybean protein, hemoglobin and bovine serum albumin. Optimal pH and temperature for reaction of the purified enzyme were 9.0 and $50^{\circ}C$, respectively. The enzyme was stable at pH 4.0-11.0, below $40^{\circ}C$, and the activity was not stimulated by metal ions. 1mM phenylmethylsulfonyl fluoride inhibited the enzyme activity by 98.5%. It means that the enzyme is one of serine protease.

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Preceding Research for Estimating the Maximal Fat Oxidation Point through Heart Rate and Heart Rate Variability (심박 및 심박변화를 통한 최대 지방 연소 시점의 추정)

  • Sim, Myeong-Heon;Kim, Min-Yong;Yoon, Chan-Sol;Chung, Joo-Hong;Noh, Yeon-Sik;Park, Sung-Bin;Yoon, Hyung-Ro
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.61 no.9
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    • pp.1340-1349
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    • 2012
  • Increasing the oxidation of fat through exercise is the recommendable method for weight control. Preceding researches have proposed increase in the usage of fat during exercise in stabilized state and under maximum exertion through aerobic training. However, such researches require additional equipment for gas analysis in order to measure the caloric value or gas exchange of subjects during exercise. Such equipments become highly restrictive for those exercise and cause substantially higher cost. According to this, we have presented the method of estimating the maximal fat oxidation point through changes in LF & HF which reflects changes in heart rate and the autonomic nervous system in order to induce exercise for a less restrictive and efficient fat oxidation than existing methods. We have conducted exercise stress test on subject with similar exercise abilities, and have detected the changes in heart rate and changes in LF & HF by measuring changes in fat oxidation and measuring ECG signals at the same time through a gas analyzer. Changes in heart rate and HRV of the subjects during exercising was detected through only the electrocardiographic signals from exercising and detected the point of maximum fat oxidation that differs from person to person. The experiment was carried out 16 healthy males, and used Modified Bruce Protocol, which is one of the methods of exercise stress test methods that use treadmill. The fat oxidation amount during exercise of all the subjects showed fat oxidation of more than 4Fkcal/min in the exercise intensity from about 5 minutes to 10 minutes. The correlation between the maximal fat oxidation point obtained through gas analysis and the point when 60% starts to be relevant in the range from -0.01 to 0.01 seconds for values of R-R interval from changes in heart rate had correlation coefficients of 0.855 in Kendall's method and in Spearman's rho, it showed significant results of it being p<0.01 with 0.950, respectively. Furthermore, in the changes in LF & HF, we have determined the point where the normalized area value starts to become the same as the maximal fat oxidation point, and the correlation here showed 0.620 in Kendall and 0.780 in Spearma of which both showed significant results as p<0.01.

Adsorption Characteristic of Brownish Dark Colored Compounds from the Hot Water Extract of Auricularia auricula Fruit Body (흑목이 버섯 자실체의 열수추출물로부터 흑갈색 색소 성분의 흡착 특성)

  • Kim, Hyeon-Min;Hur, Won;Lim, Kun Bin;Lee, Shin-Young
    • Food Engineering Progress
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    • v.13 no.2
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    • pp.138-146
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    • 2009
  • The crude polysaccharide fraction from fruit body of Auricularia auricula were obtained by using hot water extraction and ethanol precipitation. As the crude polysaccharide fraction contained the brownish dark colored compounds, the adsorption study of pigments from the crude polysaccharide using activated carbon was carried out. The pigment compounds showed an absorption characteristic with $\lambda_{max}$ of 230 nm and the absorbance at 230 nm was taken as color intensity. Adsorption capacity of pigment depended on increase of the activated carbon to sample loading ratio. The adsorption capacity increased with increase of pH and temperature in the pH range of 3.0-7.0 and temperature range of 25-40$^{\circ}C$, but decreased in the temperature range of 40-70$^{\circ}C$. The optimum capacity was obtained at addition of 16.7 mg activated carbon per mL sample solution (concentration = 3 mg/mL) at pH of 7.0 and temperature of 40$^{\circ}C$. Treatment for 10 min was sufficient to achieve the 80% decolorization and 1.25 fold purification of polysaccharide. Langmuir isotherm and pseudo second-order kinetic model provided the best fitting for adsorption of the brownish dark colored compounds onto powdered active carbon. The activation energies of adsorption from the Langmuir isotherm parameter in the ranges of 25-40$^{\circ}C$ and 40-70$^{\circ}C$ was -2.54 and 4.38 kcal/g, respectively. The results of low activation energy also indicated that the adsorption process was a physical adsorption which was controlled by diffnsion.

Evaluating Physical Characteristics of Raindrop in Anseong, Gyeonggi Province (강우입자의 물리적 특성평가: 경기도 안성시 지역을 사례로)

  • KIM, Jin Kwan;YANG, Dong Yoon;KIM, Min Seok
    • Journal of The Geomorphological Association of Korea
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    • v.17 no.1
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    • pp.49-57
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    • 2010
  • To evaluate physical characteristics of open rainfall in Korea, terminal velocity of raindrop and drop size distributions (DSD) were continuously measured using by laser-optical disdrometer around Gosam reservoir, Anseong-si, Gyeonggi-do during three rainfall events from 2008 to 2009. The relationships between kinetic energies (KE, Jm-2mm-1; KER, Jm-2h-1) and rainfall intensity were obtained, respectively. Moreover, we compared the rainfall intensity from a disdrometer with the rainfall intensity from a tipping bucket raingauge to transform the kinetic energy of rainfall using the data from a tipping bucket raingauge. Therefore, the established relationships between kinetic energies (KE and KER) and rainfall intensity could be a useful model to consider the kinetic energy of raindrop using the rainfall intensity below 40mmh-1 of max 5-min rainfall intensity in the middle of South Korea. However, to better examine the relationship between kinetic energy and rainfall intensity, further measurement will be required.

Studies on the Natural Distribution and Ecology of Ilex cornuta Lindley et Pax. in Korea (호랑가시나무의 천연분포(天然分布)와 군낙생태(群落生態)에 관한 연구(研究))

  • Lee, Jeong Seok
    • Journal of Korean Society of Forest Science
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    • v.62 no.1
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    • pp.24-42
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    • 1983
  • To develop Ilex cornuta which grow naturally in the southwest seaside district as new ornamental tree, the author chose I. cornuta growing in the four natural communities and those cultivated in Kwangju city as a sample, and investigated its ecology, morphology and characteristics. The results obtained was summarized as follows; 1) The natural distribution of I. cornuta marks $35^{\circ}$43'N and $126^{\circ}$44'E in the southwestern part of Korea and $33^{\circ}$20'N and $126^{\circ}$15'E in Jejoo island. This area has the following necessary conditions for Ilex cornuta: the annual average temperature is above $12^{\circ}C$, the coldness index below $-12.7^{\circ}C$, annual average relative humidity 75-80%, and the number of snow-covering days is 20-25 days, situated within 20km of from coastline and within, 100m above sea level and mainly at the foot of the mountain facing the southeast. 2) The vegetation in I. cornuta community can be divided that upper layer is composed of Pinus thunbergii and P. densiflora, middle layer of Eurya japonica var. montana, Ilex cornuta and Vaccinium bracteatum, and the ground vegetation is composed of Carex lanceolata and Arundinella hirta var. ciliare. The community has high species diversity which indicates it is at the stage of development. Although I. cornuta is a species of the southern type of temperate zone where coniferous tree or broad leaved, evergreen trees grow together, it occasionally grows in the subtropical zone. 3) Parent rock is gneiss or rhyolite etc., and soil is acidic (about pH 4.5-5.0) and the content of available phosphorus is low. 4) At maturity, the height growth averaged $10.48{\pm}0.23cm$ a year and the diameter growth 0.43 cm a year, and the annual ring was not clear. Mean leaf-number was 11.34. There are a significant positive correlation between twig-elongation and leaf-number. 5) One-year-old seedling grows up to 10.66 cm (max. 18.2 cm, min. 4.0 cm) in shoot-height, with its leaf number 12.1 (max. 18, min), its basal diameter 2.24 mm (max. 4.0 mm, min. 1.0 mm) and shows rhythmical growth in high temperature period. There were significant positive correlations between stalk-height and leaf-number, between stalk-height and basal-diameter, and between number and basal diameter. 6) The flowering time ranged from the end of April to the beginning of May, and the flower has tetra-merouscorella and corymb of yellowish green. It has a bisexual flower and dioecism with a sexual ratio 1:1. 7) The fruit, after fertilization, grows 0.87 cm long (0.61-1.31 cm) and 0.8 cm wide (0.62-1.05 cm) by the beginning of May. Fruits begin to turn red and continue to ripen until the end of October or the beginning of November and remain unfading until the end of following May. With the partial change in color of dark-brown at the beginning of the June fruits begin to fall, bur some remain even after three years. 8) The seed acquision ratio is 24.7% by weight, and the number of grains per fruit averages 3.9 and the seed weight per liter is 114.2 gram, while the average weight of 1,000 seeds is 24.56 grams. 9) Seeds after complete removal of sarcocarp, were buried under ground in a fixed temperature and humidity and they began to develop root in October, a year later and germinated in the next April. Under sunlight or drought, however, the dormant state may be continued.

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Development of Dual Reporter System of Mutant Dopamine 2 Receptor ($D_2R$) and Sodium Iodide Symporter (NIS) Transgenes (변이 도파민 2 수용체와 나트륨 옥소 공동 수송체 이입유전자의 이중 리포터시스템 개발)

  • Hwang, Do-Won;Lee, Dong-Soo;Kang, Joo-Hyun;Chang, Young-Soo;Kim, Yun-Hui;Jeong, Jae-Min;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.4
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    • pp.294-299
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    • 2004
  • Purpose: Both human NIS and mutant $D_2R$ transgenes are proposed as reporting system in transplanted cell tracking. Using hepatoma cell lines, we constructed a dual reporter system containing human sodium-iodide symporter (hNIS) and dopamine 2 receptor ($D_2R$) and compared its characteristics. Materials and Methods: The recombinant plasmid ($pIRES-hNIS/D_2R$) was constructed with IRES (internal ribosome entry site) under control of the CMV promoter $pIRES-hNIS/D_2R$ was transfected to human hepatoma SK-Hep1 cell line with lipofectamine. HEP-ND ($SK-Hep1-hNIS/D_2R$) cells stably expressing hNIS and $D_2R$ was established by selection with G418 for two weeks. RT-PCR was performed to investigate the expression of both hNIS and $D_2R$ genes. The expressions of hNIS and $D_2R$ were measured by $^{125}I$ uptake assays and receptor binding assays. Specific binding of $D_2R$ to $[^3H]spiperone$ was verified by Scatchard plot with (+) butaclamol as a specific inhibitor. $K_d\;and\;B_{max}$ values were estimated. The correlation between hNIS and $D_2R$ expression was compared by using each clone. Results: Similar quantities of hNIS and $D_2R$ genes were expressed on HEP-ND as RT-PCR assays. HEP-ND cells showed 30 to 40 fold higher radioiodine uptakes than those of parental SK-Hep1 cells. $^{125}I$ uptake in HEP-ND cells was completely inhibited by $KClO_4$, a NIS inhibitor Specific binding to HEP-ND cells was saturable and the $K_d\;and\;B_{max}$ values for HEP-ND cells were 2.92 nM, 745.25 fmol/mg protein and 2.91nM, 1323 fmole/mg protein in two clones, respectively. The radioiodine uptake by hNIS activity and $D_2R$ binding was highly correlated. Conclusion: We developed a dual positron and gamma imaging reporter system of hNIS and $D_2R$ in a stably transfected cell line. We expect that $D_2R$ and hNIS genes can complement mutually as a nuclear reporting system or that $D_2R$ can be used as reporter gene when hNIS gene were used as a treatment gene.

Characteristic Evaluation of Optically Stimulated Luminescent Dosimeter (OSLD) for Dosimetry (광유도발광선량계(Optically Stimulated Luminescent Dosimeter)의 선량 특성에 관한 고찰)

  • Kim, Jeong-Mi;Jeon, Su-Dong;Back, Geum-Mun;Jo, Young-Pil;Yun, Hwa-Ryong;Kwon, Kyung-Tae
    • The Journal of Korean Society for Radiation Therapy
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    • v.22 no.2
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    • pp.123-129
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    • 2010
  • Purpose: The purpose of this study was to evaluate dosimetric characteristics of Optically stimulated luminescent dosimeters (OSLD) for dosimetry Materials and Methods: InLight/OSL $NanoDot^{TM}$ dosimeters was used including $Inlight^{TM}MicroStar$ Reader, Solid Water Phantom, and Linear accelerator ($TRYLOGY^{(R)}$) OSLDs were placed at a Dmax in a solid water phantom and were irradiated with 100 cGy of 6 MV X-rays. Most irradiations were carried out using an SSD set up 100 cm, $10{\times}10\;cm^2$ field and 300 MU/min. The time dependence were measured at 10 minute intervals. The dose dependence were measured from 50 cGy to 600 cGy. The energy dependence was measured for nominal photon beam energies of 6, 15 MV and electron beam energies of 4-20 MeV. The dose rate dependence were also measured for dose rates of 100-1,000 MU/min. Finally, the PDD was measured by OSLDs and Ion-chamber. Results: The reproducibility of OSLD according to the Time flow was evaluated within ${\pm}2.5%$. The result of Linearity of OSLD, the dose was increased linearly up to about the 300 cGy and increased supralinearly above the 300 cGy. Energy and dose rate dependence of the response of OSL detectors were evaluated within ${\pm}2%$ and ${\pm}3%$. $PDD_{10}$ and PDD20 which were measured by OSLD was 66.7%, 38.4% and $PDD_{10}$ and $PDD_{20}$ which were measured by Ion-chamber was 66.6%, 38.3% Conclusion: As a result of analyzing characteration of OSLD, OSLD was evaluated within ${\pm}3%$ according to the change of the time, enregy and dose rate. The $PDD_{10}$ and $PDD_{20}$ are measured by OSLD and ion-chamber were evaluated within 0.3%. The OSL response is linear with a dose in the range 50~300 cGy. It was possible to repeat measurement many times and progress of the measurement of reading is easy. So the stability of the system and linear dose response relationship make it a good for dosimetry.

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Study of Factors Controlling Exposure Dose and Image Quality of C-arm in Operation Room according to Detector Size of It (Mainly L-Spine AP Study) (수술 중 C-Arm Neutral AP 검사 시 조절인자에 따른 피폭선량 및 화질비교(L-Spine AP검사를 기준으로))

  • CHOI, Sung-Hyun;JO, Hwang-Woo;Dong, Kyung-Rae;Chung, Woon-Kwan;Choi, Eun-Jin;Song, Ha-jin
    • Journal of Radiation Industry
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    • v.9 no.2
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    • pp.85-90
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    • 2015
  • Purpose: Time of operation has been reduced and accuracy of operation has been improved since C-arm, which offer real-time image of patient, was introduced in operation room. However, because of the contamination of patient, C-arm could not be used more appropriately. Therefore, this study is to know factors of controlling exposure dose, image quality and the exposed dose of health professional in operation room. Materials and methods: Height of Wilson frame (bed for operation) was fixed at 130 cm. Then, Model 76-2 Phantom, which was set by assembling manual of Fluke Company, was set on the bed. Head/Spine Fluoroscopy AEC mode was set for exposure condition. According to detector size of C-arm, the absorbed dose per min was measured in the 7 steps OFD (cm) from 10 cm to 40 cm (10, 15, 20, 25, 30, 35, 40 cm). In each step of OFD, the absorbed dose per min of same diameter of collimation was measured. Moreover, using Nero MAX Model 8000, exposure dose per min was measured according to 3 step of distance from detector (20 cm, 60 cm, 100 cm). Finally, resolution was measured by CDRH Disc Phantom and magnification of each OFD was measured by aluminum stick bar. Result: According to detector size of C-arm, difference of absorbed dose shows that the dose of 20 cm OFD is 1.750 times higher than the dose of 40 cm OFD. It means that the C-arm, which has smaller size of detector, shows the bigger difference of absorbed dose per min (p<0.05). In the difference of absorbed dose in the same step of OFD (from 20 cm to 40 cm), the absorbed dose of 9 inch detect or C-arm was 1.370 times higher than 12 inch' s (p<0.05). When OFD was set to 20 cm OFD, the absorbed dose of non-collimation case was approximately 0.816 times lower than the absorbed dose of collimation cases (p<0.05). When the distance was 20 cm from detector, exposed does includes first-ray and scatter-ray. When the distance was 60 cm and 100 cm from detector, exposed does includes just scatter-ray. So, there was the 2.200 times difference of absorbed does. Finally, when OFD was increased, spatial resolution was 4 to 5 step was increased. However, low contrast resolution was not relative. Moreover, there was 1.363 times difference of magnification (p<0.05). Conclusion: When C-Arm is used, avoiding contamination of patient is more important factor than reducing exposed dose of health professional in operation room. Just controlling exposure time is just way to reduce the exposed does of workers. However, in the case, non-probability influence could be occurred. Therefore, this study proved that the exposed dose will be reduced if the factors such as using small detector size of C-arm, setting OFD from 20 cm to 25 cm and non-collimating. Moreover, dose management of C-arm in the non-interesting area will be considered additionally.

Evaluation of Dose Change by Using the Deformable Image Registration (DIR) on the Intensity Modulated Radiation Therapy (IMRT) with Glottis Cancer (성문암 세기조절 방사선치료에서 변형영상정합을 이용한 선량변화 평가)

  • Kim, Woo Chul;Min, Chul Kee;Lee, Suk;Choi, Sang Hyoun;Cho, Kwang Hwan;Jung, Jae Hong;Kim, Eun Seog;Yeo, Seung-Gu;Kwon, Soo-Il;Lee, Kil-Dong
    • Progress in Medical Physics
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    • v.25 no.3
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    • pp.167-175
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    • 2014
  • The purpose of this study is to evaluate the variation of the dose which is delivered to the patients with glottis cancer under IMRT (intensity modulated radiation therapy) by using the 3D registration with CBCT (cone beam CT) images and the DIR (deformable image registration) techniques. The CBCT images which were obtained at a one-week interval were reconstructed by using B-spline algorithm in DIR system, and doses were recalculated based on the newly obtained CBCT images. The dose distributions to the tumor and the critical organs were compared with reference. For the change of volume depending on weight at 3 to 5 weeks, there was increased of 1.38~2.04 kg on average. For the body surface depending on weight, there was decreased of 2.1 mm. The dose with transmitted to the carotid since three weeks was increased compared be more than 8.76% planned, and the thyroid gland was decreased to 26.4%. For the physical evaluation factors of the tumor, PITV, TCI, rDHI, mDHI, and CN were decreased to 4.32%, 5.78%, 44.54%, 12.32%, and 7.11%, respectively. Moreover, $D_{max}$, $D_{mean}$, $V_{67.50}$, and $D_{95}$ for PTV were increased or decreased to 2.99%, 1.52%, 5.78%, and 11.94%, respectively. Although there was no change of volume depending on weight, the change of body types occurred, and IMRT with the narrow composure margin sensitively responded to such a changing. For the glottis IMRT, the patient's weight changes should be observed and recorded to evaluate the actual dose distribution by using the DIR techniques, and more the adaptive treatment planning during the treatment course is needed to deliver the accurate dose to the patients.

Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars (국내 콩(Glycine max) 품종 형질전환 초기조건 확립)

  • Lee Ki-Jung;Seo Jen-Kyung;Lee Hye-Young;Jeon Eun-Hee;Shin Sang-Hyun;Lee Jai-Heon;Kim Doh-Hoon;Ko Jong-Min;Hahn Won Young;Baek In-Youl;Oh Boung-Jun;Chung Young-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.289-296
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    • 2006
  • In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.