• Title/Summary/Keyword: co-transformation

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Bioequivalence of Broadcef Capsule to Cefradine Yuhan Capsule (Cephradine 500 mg) (유한세프라딘 캅셀(세프라딘 500 mg)에 대한 브로드세프 캅셀의 생물학적 동등성)

  • Cho, Hea-Young;Lee, Suk;Kang, Hyun-Ah;Oh, In-Joon;Lim, Dong-Koo;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.32 no.3
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    • pp.215-221
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    • 2002
  • Cephradine is a first generation cephalosporin and has broad spectrum antibacterial activity against gram-positive and gram-negative microorganisms, through inhibition of bacterial cell wall synthesis. Cephradine is useful for treatment of infections of the urinary and respiratory tract, skin and soft tissues. The purpose of the present study was to evaluate the bioequivalence of two cephradine capsules, Cefradine Yuhan (YuHan Corporation) and Broadcef (Ilsung Pharmaceuticals Co. Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). The cephradine release from the two cephradine capsules in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty normal male volunteers, $23.10{\pm}2.90$ years in age and $67.69{\pm}8.04\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one capsule containing 500 mg as cephradine was orally administered, blood was taken at predetermined time intervals and the concentrations of cephradine in serum were determined using HPLC method with UV detector. The dissolution profiles of two cephradine capsules were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AVC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AVC_t\;and\;C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AVC_t,\;C_{max}\;and\;T_{max}$ between two capsules based on the Cefradine Yuhan were -2.87%, -0.96% and -4.85%, respectively. There were no sequence effects between two capsules in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of 1og(0.8) to log(1.25) $(e.g.,\;log(0.93){\sim}log(1.02)\;and\;log(0.88){\sim}log(1.13)\;for \;AVC_t\;and\;C_{max},\;respectively)$. The 90% confidence interval using untransformed data was within ${\pm}20%$ $(e.g., \;-17.54{\sim}7.78\;for\;T_{max})$. All parameters met the criteria of KFDA guideline for bioequivalence, indicating that Broadcef capsule is bioequivalent to Cefradine Yuhan capsule.

Bioequivalence of Flucona Capsule to Diflucan Capsule (Fluconazole 50 mg) (디푸루칸 캅셀(플루코나졸 50 mg)에 대한 플루코나 캅셀의 생물학적 동등성)

  • Cho, Hea-Young;Kang, Hyun-Ah;Lee, Suk;Oh, In-Joon;Lim, Dong-Koo;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.33 no.2
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    • pp.135-140
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    • 2003
  • Fluconazole is an orally active bis-triazole antifungal agent, which is used in the treatment of superficial and systemic candidiasis and in the treatment of cryptococcal infections in patients with the acquired immuno deficiency syndrome (AIDS). The purpose of the present study was to evaluate the bioequivalence of two fluconazole capsules, Diflucan (Pfizer Pharmaceuticals Korea Inc.) and Flucona (Korean Drug Pharmaceuticals Co., Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). The fluconazole release from the two fluconazole capsules in vitro was tested using KP VII Apparatus II method at 0.1 M hydrochloride dissolution media. Twenty normal male volunteers, $23.60{\pm}1.88$ years in age and $63.57{\pm}6.17\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After three capsules containing 50 mg as fluconazole was orally administered, blood was taken at predetermined time intervals and the concentrations of fluconazole in serum were determined using HPLC method with UV detector. The dissolution profiles of two fluconazole capsules were very similar at 0.1 M hydrochloride dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$ and $C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AUC_t,\;C_{max}\;and\;T_{max}$ between two capsules based on the Diflucan were 4.96%, 5.65% and -13.76%, respectively. There were no sequence effects between two capsules in these parameter. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) $(e.g.,\;log(1.01){\sim}log(1.08)\;and\;log(1.00){\sim}log(1.12)\;for\;AUC_t\;and\;C_{max},\;respectively)$, indicating that Flucona capsule is bioequivalent to Diflucan capsule.

Effect of Epidermal Growth Factor (EGF) on Meiotic Maturation and Pronuclear Formation of Porcine Oocytes Produced In Vitro

  • Song S. H.;Kim J. G.;Song H. J.;Kumar B. Mohana;Cho S. R.;Choe C. Y.;Choi S. H.;Rho G. J.;Choe S. Y.
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.127-131
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    • 2005
  • The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

Polymorphisms and Functional Analysis of the Intact Human Papillomavirus16 E2 Gene

  • Ekalaksananan, Tipaya;Jungpol, Watcharapol;Prasitthimay, Chuthamas;Wongjampa, Weerayut;Kongyingyoes, Bunkerd;Pientong, Chamsai
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10255-10262
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    • 2015
  • High risk human papillomavirus (HR-HPV) E2 proteins play roles in transcriptional regulation and are commonly functionally disrupted when the HPV genome integrates into host chromosomes. Some 15-40% of cancer cases, however, contain an intact E2 gene or episomal HPV. In these cases, polymorphism of the E2 gene might be involved. This study aimed to determine polymorphisms of the E2 gene in episomal HPV16 detected in high grade squamous intraepithelial lesions and squamous cell carcinomas and altered functions compared to the E2 prototype. The E2 gene was amplified and sequenced. Two expression vectors containing E2 gene polymorphisms were constructed and transfected in SiHa and C33A cells, then E6 gene as well as Il-10 and TNF-${\alpha}$ expression was determined by quantitative RT-PCR. Expression vectors and reporter vectors containing the HPV16 long control region (LCR) were co-transfected and transcriptional activity was determined. The results showed that a total of 32 nucleotides and 23 amino acids were changed in all 20 cases of study, found in the transactivation (TA) domain, hinge (H) region and DNA binding (DB) domain with 14, 5 and 13 nucleotide positions. They mostly caused amino acid change. The expressing vectors containing different E2 gene polymorphisms showed E6 mRNA suppression, TNF-${\alpha}$ mRNA suppression and IL-10 induction but no statistically significant differences when compared to the E2 prototype. Moreover, promoter activity in HPV16 LCR was not affected by E2 protein with different gene polymorphisms, in contrast to nucleotide variations in LCR that showed an effect on transcription activity. These results demonstrated that E2 gene polymorphisms of episomal HPV16 did not affect transcriptional regulation and suggested that nucleotide variation as well as epigenetic modification of the LCR might play a role in inducing malignant transformation of cells containing episomal HPV16.

Evaluation of a Traffic Noise Predictive Model for an Active Noise Cancellation (ANC) System (능동형 소음저감 기법을 위한 도로교통소음 예측 모형 평가 연구)

  • An, Deok Soon;Mun, Sung Ho;An, Oh Seong;Kim, Do Wan
    • International Journal of Highway Engineering
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    • v.17 no.6
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    • pp.11-18
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    • 2015
  • PURPOSES : The purpose of this thesis is to evaluate the effectiveness of an active noise cancellation (ANC) system in reducing the traffic noise level against frequencies from the predictive model developed by previous research. The predictive model is based on ISO 9613-2 standards using the Noble close proximity (NCPX) method and the pass-by method. This means that the use of these standards is a powerful tool for analyzing the traffic noise level because of the strengths of these methods. Traffic noise analysis was performed based on digital signal processing (DSP) for detecting traffic noise with the pass-by method at the test site. METHODS : There are several analysis methods, which are generally divided into three different types, available to evaluate traffic noise predictive models. The first method uses the classification standard of 12 vehicle types. The second method is based on a standard of four vehicle types. The third method is founded on 5 types of vehicles, which are different from the types used by the second method. This means that the second method not only consolidates 12 vehicle types into only four types, but also that the results of the noise analysis of the total traffic volume are reflected in a comparison analysis of the three types of methods. The constant percent bandwidth (CPB) analysis was used to identify the properties of different frequencies in the frequency analysis. A-weighting was applied to the DSP and to the transformation process from analog to digital signal. The root mean squared error (RMSE) was applied to compare and evaluate the predictive model results of the three analysis methods. RESULTS : The result derived from the third method, based on the classification standard of 5 vehicle types, shows the smallest values of RMSE and max and min error. However, it does not have the reduction properties of a predictive model. To evaluate the predictive model of an ANC system, a reduction analysis of the total sound pressure level (TSPL), dB(A), was conducted. As a result, the analysis based on the third method has the smallest value of RMSE and max error. The effect of traffic noise reduction was the greatest value of the types of analysis in this research. CONCLUSIONS : From the results of the error analysis, the application method for categorizing vehicle types related to the 12-vehicle classification based on previous research is appropriate to the ANC system. However, the performance of a predictive model on an ANC system is up to a value of traffic noise reduction. By the same token, the most appropriate method that influences the maximum reduction effect is found in the third method of traffic analysis. This method has a value of traffic noise reduction of 31.28 dB(A). In conclusion, research for detecting the friction noise between a tire and the road surface for the 12 vehicle types needs to be conducted to authentically demonstrate an ANC system in the Republic of Korea.

Field Performance and Morphological Characterization of Transgenic Codonopsis lanceolata Expressing $\gamma-TMT$ Gene.

  • Ghimire, Bimal Kumar;Li, Cheng Hao;Kil, Hyun-Young;Kim, Na-Young;Lim, Jung-Dae;Kim, Jae-Kwang;Kim, Myong-Jo;Chung, Ill-Min;Lee, Sun-Joo;Eom, Seok-Hyun;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.5
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    • pp.339-345
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    • 2007
  • Field performance and morphological characterization was conducted on seven transgenic lines of Codonopsis lanceolata expressing ${\gamma}-TMT$ gene. The shoots were obtained from leaf explants after co-cultivation with Agrobacterium tume-faciens strain LBA 4404 harboring a binary vector pYBI 121 that carried genes encoding ${\gamma}-Tocopherol$ methyltransferase gene (${\gamma}-TMT$) and a neomycin phosphotransferase II gene (npt II) for kanamycin resistance. The transgenic plants were transferred to a green house for acclimation. Integration of T-DNA into the $T_0\;and\;T_1$ generation of transgenic Codonopsis lanceolata genome was confirmed by the polymerase chain reaction and southern blot analysis. The progenies of transgenic plants showed phenotypic differences within the different lines and with relative to control plants. When grown in field, the transgenic plants in general exhibited increased fertility, significant improvement in the shoot weight, root weight, shoot height and rachis length with relation to the control plants. However, all seven independently derived transgenic lines produced normal flower with respect to its shape, size, color and seeds number at its maturity. Indicating that the addition of a selectable marker gene in the plant genome does not effect on seed germination and agronomic performance of transgenic Codonopsis lanceolata. $T_1$ progenies of these plants were obtained and evaluated together with control plant in a field experiment. Overall, the agronomic performance of $T_1$ progenies of transgenic Codonopsis lanceolata showed superior to that of the seed derived non-transgenic plant. In this study, we report on the morphological variation and agronomic performance of transgenic Codonopsis lanceolata developed by Agrobacterium transformation.

Thermal Stable Ni-silicide Utilizing Pd Stacked Layer for nano-scale CMOSFETs (나노급 CMOSFET을 위한 Pd 적층구조를 갖는 열안정 높은 Ni-silicide)

  • Yu, Ji-Won;Zhang, Ying-Ying;Park, Kee-Young;Li, Shi-Guang;Zhong, Zhun;Jung, Soon-Yen;Yim, Kyoung-Yean;Lee, Ga-Won;Wang, Jin-Suk;Lee, Hi-Deok
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.10-10
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    • 2008
  • Silicide is inevitable for CMOSFETs to reduce RC delay by reducing the sheet resistance of gate and source/drain regions. Ni-silicide is a promising material which can be used for the 65nm CMOS technologies. Ni-silicide was proposed in order to make up for the weak points of Co-silicide and Ti-silicide, such as the high consumption of silicon and the line width limitation. Low resistivity NiSi can be formed at low temperature ($\sim500^{\circ}C$) with only one-step heat treat. Ni silicide also has less dependence of sheet resistance on line width and less consumption of silicon because of low resistivity NiSi phase. However, the low thermal stability of the Ni-silicide is a major problem for the post process implementation, such as metalization or ILD(inter layer dielectric) process, that is, it is crucial to prevent both the agglomeration of mono-silicide and its transformation into $NiSi_2$. To solve the thermal immune problem of Ni-silicide, various studies, such as capping layer and inter layer, have been worked. In this paper, the Ni-silicide utilizing Pd stacked layer (Pd/Ni/TiN) was studied for highly thermal immune nano-scale CMOSFETs technology. The proposed structure was compared with NiITiN structure and showed much better thermal stability than Ni/TiN.

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Development of high tryptophan GM rice and its transcriptome analysis (고 함량 트립토판 생산 GM 벼 개발 및 전사체 분석)

  • Jung, Yu Jin;Nogoy, Franz Marielle;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.186-195
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    • 2015
  • Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

Control Indian meal moth Plodia interpunctella by gas treatment

  • Han, Gyung Deok;Kwon, Hyeok;Jin, Hyun Jung;Kum, Ho Jung;Kim, Bo Hwan;Kim, Wook
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.45-45
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    • 2017
  • The Indian meal moth, Plodia interpunctella, is one of the most important pests of stored food in the food processing industry worldwide. To control the Indian meal moth, methyl bromide, phosphine, high carbon dioxide, sulfuryl fluoride and plant essential oil fumigation have been considered. However, these treatments have disadvantages. For example, depleting the ozone layer, showing resistance in insect, low control efficacy or need high cost for treatment. Chlorine dioxide ($ClO_2$) is strong disinfectant and insecticide. The gas caused a malfunction in enzymes. The oxidative stress induced by $ClO_2$ gas treatment damaged to a physiological system and all life stages of P. interpunctella. The gaseous $ClO_2$ is a convincing alternative to methyl bromide for controlling P. interpunctella. The gaseous $ClO_2$ was generated by a chlorine dioxide generator (PurgoFarm Co., Ltd., Hwasung, Korea). It generated highly pure $ClO_2$ gas and the gas blown out through a vent into a test chamber. Gas entry to the chamber was automatically controlled and monitored by a PortaSene II gas leak detector (Analytical Technology, Collegeville, PA, USA). The properly prepared eggs, larvae, pupae, and adults of P. interpunctella were used in this experiment. Data were analyzed using SAS 9.4. Percentage data were statistically analyzed after arcsine-root transformation. Analysis of variance was performed using general linear model, and means were separated by the least significant difference test at P < 0.05. Fumigation is an effective management technique for controlling all stages of P. interpunctella. We found that $ClO_2$ gas treatment directly effects on egg, larvae, pupae and adults of P. interpunctella. The gas treatment with proper concentration for over a day achieved 100 % mortality in all stages of P. interpunctella and short time treatment or low concentration gas treatment results showed that the egg hatchability, pupation rate, and adult emergency rate were lowered compare with untreated control. Also, abnormal pupae or adult rate were increased. Gaseous $ClO_2$ treatment induced insecticidal reactive oxygen species (ROS), and it resulted in fatal oxidative stress in P. interpunctella. Taken together, these results showed that exposure proper concentration and time of the gas control all stages of P. interpunctella by inducing fatal oxidative stress. Further studies will be required to apply the gas treatment under real-world condition and to understanding physiological reaction in P. interpunctella caused by oxidative stress.

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Development of transgenic cucumbers expressing Arabidopsis Nit gene (애기장대 Nit유전자 발현 오이 형질전환체 개발)

  • Jang, Hyun A;Lim, Ka Min;Kim, Hyun A;Park, Yeon-Il;Kwon, Suk Yoon;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.198-202
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    • 2013
  • To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.