• Title/Summary/Keyword: Suspension method

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Improvement of Proliferation Capacity of Non-adapted CHO Cells Subcultured Using Serum Free Media in Long-term Culture (무혈청 배지에서 계대배양한 비적응 CHO(Chinese Hamster Ovary) 세포의 증식력 개선에 관한 연구)

  • Lee, Seung-Sun;Lee, Jin-Sung;Byun, Soon-Hyu;Park, Hong-Woo;Choe, Tae-Boo
    • KSBB Journal
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    • v.21 no.4
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    • pp.248-254
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    • 2006
  • Animal cell culture industry has a large market and an exponential growth rate among biological industry field. Chines hamster ovary(CHO) cells are the most widely used cell lines for recombinant protein production. They can avoid infection from polio, herpes, hepatitis B, HIV, measles, adenovirus and etc. Moreover it is easy to transfection recombinant genes and possible to suspension culture. Serum free media is one of the most important factor of protein production. Because serum has problems. Serum is not defined the contents until now, it has a number of proteins, lipids, carbohydrates and unknown molecules that cause of risk involve in infection and high cost of product purification. CHO cell line cultured using serum free media were the basis of a very successful method to produce(glyco-)protein in mammalian cells, which are then used as pharmaceutical products. Also, the low protein content of the developed medium facilitates downstream processing and product purification. But non-adapted CHO cells have a limit of proliferation cultured using serum free media and it takes very long time to adapt non-adapted cells to serum free media. There are a number of causes of a limit of proliferation using serum free media. Absence of growth factors and growth stimulating molecules is a major factor of the reasons. It makes growth signals and moves cell cycle. And increase of cellular stress is another reason. It induces increase of intraceullar ROS concentration. The purpose of this study is about improvement of proliferation capacity of non-adapted CHO cells cultured using serum free media without adaptation process.

Carbon nanotube field emission display

  • Chil, Won-Bong;Kim, Jong-Min
    • Electrical & Electronic Materials
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    • v.12 no.7
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    • pp.7-11
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    • 1999
  • Fully sealed field emission display in size of 4.5 inch has been fabricated using single-wall carbon nanotubes-organic vehicle com-posite. The fabricated display were fully scalable at low temperature below 415$^{\circ}C$ and CNTs were vertically aligned using paste squeeze and surface rubbing techniques. The turn-on fields of 1V/${\mu}{\textrm}{m}$ and field emis-sion current of 1.5mA at 3V/${\mu}{\textrm}{m}$ (J=90${\mu}{\textrm}{m}$/$\textrm{cm}^2$)were observed. Brightness of 1800cd/$m^2$ at 3.7V/${\mu}{\textrm}{m}$ was observed on the entire area of 4.5-inch panel from the green phosphor-ITO glass. The fluctuation of the current was found to be about 7% over a 4.5-inch cath-ode area. This reliable result enables us to produce large area full-color flat panel dis-play in the near future. Carbon nanotubes (CNTs) have attracted much attention because of their unique elec-trical properties and their potential applica-tions [1, 2]. Large aspect ratio of CNTs together with high chemical stability. ther-mal conductivity, and high mechanical strength are advantageous for applications to the field emitter [3]. Several results have been reported on the field emissions from multi-walled nanotubes (MWNTs) and single-walled nanotubes (SWNTs) grown from arc discharge [4, 5]. De Heer et al. have reported the field emission from nan-otubes aligned by the suspension-filtering method. This approach is too difficult to be fully adopted in integration process. Recently, there have been efforts to make applications to field emission devices using nanotubes. Saito et al. demonstrated a car-bon nanotube-based lamp, which was oper-ated at high voltage (10KV) [8]. Aproto-type diode structure was tested by the size of 100mm $\times$ 10mm in vacuum chamber [9]. the difficulties arise from the arrangement of vertically aligned nanotubes after the growth. Recently vertically aligned carbon nanotubes have been synthesized using plasma-enhanced chemical vapor deposition(CVD) [6, 7]. Yet, control of a large area synthesis is still not easily accessible with such approaches. Here we report integra-tion processes of fully sealed 4.5-inch CNT-field emission displays (FEDs). Low turn-on voltage with high brightness, and stabili-ty clearly demonstrate the potential applica-bility of carbon nanotubes to full color dis-plays in near future. For flat panel display in a large area, car-bon nanotubes-based field emitters were fabricated by using nanotubes-organic vehi-cles. The purified SWNTs, which were syn-thesized by dc arc discharge, were dispersed in iso propyl alcohol, and then mixed with on organic binder. The paste of well-dis-persed carbon nanotubes was squeezed onto the metal-patterned sodalime glass throuhg the metal mesh of 20${\mu}{\textrm}{m}$ in size and subse-quently heat-treated in order to remove the organic binder. The insulating spacers in thickness of 200${\mu}{\textrm}{m}$ are inserted between the lower and upper glasses. The Y\ulcornerO\ulcornerS:Eu, ZnS:Cu, Al, and ZnS:Ag, Cl, phosphors are electrically deposited on the upper glass for red, green, and blue colors, respectively. The typical sizes of each phosphor are 2~3 micron. The assembled structure was sealed in an atmosphere of highly purified Ar gas by means of a glass frit. The display plate was evacuated down to the pressure level of 1$\times$10\ulcorner Torr. Three non-evaporable getters of Ti-Zr-V-Fe were activated during the final heat-exhausting procedure. Finally, the active area of 4.5-inch panel with fully sealed carbon nanotubes was pro-duced. Emission currents were character-ized by the DC-mode and pulse-modulating mode at the voltage up to 800 volts. The brightness of field emission was measured by the Luminance calorimeter (BM-7, Topcon).

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Colony Size Distributions according to in vitro Aging in Human Skin Fibroblasts (피부 섬유모세포 노화에 따른 세포집락 크기의 분포)

  • Kim, Jun-Sang;Kim, Jae-Sung;Cho, Moon-June;Park, Jeong-Kyu;Park, Tae-Hyun
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.158-165
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    • 1999
  • Purpose : To investigate the percentage of colonies wi1h16or more cells distribution of human skin fibroblast according to in vitro aging, and to evaluate the relationship between percentage of colonies with 10 or more cells and in vivo donor age in human skin fibroblast culture. Material and Method : C1, C2, C3a, and C3b human skin fibroblast samples from three breast cancer patients were used as subjects. The C1, C2, and C3a donor were 44, 54, and 55 years old, respectively. C3a and C3b cells were isolated from the same person. Single cell suspension of skin fibroblasts was prepared with primary explant technique. One hundred cells are plated into 100m1 tissue culture flask and cultured for two weeks. The colony size was defined as colonies with 16 or more cells. The cultured cell was stained with crystal violet, and number of cells in each colony was determined with stereo microscope at $\times$10 magnification. Passage number of C1, C2, C3a and C3b skin fibroblast were 12th, 17th, and 14th, respectively. Results : Percentage of colonies with 16 or more cells of skin fibroblast samples decreased with increasing in vitro passage number. In contrast, cumulative population doublings of skin fibroblast sample increased with increasing in vitro passage number. Percentage of colonies with 16 or more cells also decreased with increasing population doublings in human skin fibroblast culture. There was strong correlation with percentage of colonies with 16 or more cells and population doublings En C3a skin fibroblast sampie. At the same point of population doublings, the percentage of colonies with 16 or more cells of the young C1 donor was higher level than the old C3a donor. Conclusion : The population doublings increased with increasing in vitro passage number but percentage of colonies with 16 or more cells decreased. The results of this study imply that percentage of colonies with 16 or more cell is useful as a indicator of in vitro human skin fibroblast aging and may estimate the in vivo donor age.

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Studies on Application Method and Safety of Macrogranule(GG) Herbicide for Remote-controlled Aerial Application (무인헬기용 Macrogranule(GG) 제초제의 안정성 및 살포방법 연구)

  • Yoon, Cheol-Su;Bae, Chang-Hyu;Lee, Sheong-Chun;Kim, Kyung-Hyun;Lee, Kye-Hwan;Cho, Tae-Kyoung;Hwang, In-Cheon
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.294-307
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    • 2011
  • This experiment carried out to confirm characteristics of macrogranule (GG) for herbicidal efficacy by using remote-controlled aerial application (RCAA) to control annual and perennial weeds in rice paddy field, application methods, and application times of formulation types. Particle of 500 g GG having average diameter of 2.5~3 mm was over 85 percent, the bulk density of the particle was $0.2\sim0.4g\;mL^{-1}$ and the particle was water floating granule. Active ingredients and external form of halosulfuronmethyl+mefenacet GG remained stable under condition of storage stability test. The disintegration time of the GG was faster as the water temperature was increased. But disintegration time was not affected by pH on the water conditions. By using hand, spoon and power applicator, drift distances of GG were 4~5m, 7 m and 10~12 m, respectively. GG showed good herbicidal efficacy and plant safety in all of the application methods such as, using hand, spoon, power applicator and RCAA. Application times of GG, GR, SC and TB by hand in $4,000m^2$ were 38 min. 4 sec, 42 min. 20 sec, 38 min. 10 sec and 21 min. 4 sec, respectively, but application time of GG by using RCAA was 1 min 32 sec. According to appearance and characteristics of formulation types, suspension concentrate (SC) and GG were possible formulation types for RCAA.

Assessment of Both Standard and Isolated Vibrio parahaemolyticus on Efficacy of Commercial Sanitizers and Disinfectants (Vibrio parahaemolyticus 표준 및 식품분리 균주에 대한 살균소독제 유효성분별 감수성 평가)

  • Kim, Il-Jin;Kim, Yong-Su;Kim, Hyung-Il;Choi, Hyun-Chul;Jeon, Dea-Hoon;Lee, Young-Ja;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.22 no.2
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    • pp.127-131
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    • 2007
  • This study evaluated the bactericidal effect of 10 sanitizers and disinfectants such as ethanol (75 and 95%), iodine (15 and 25 ppm), chlorine (100 and 200 ppm), quaternary ammonium, acid, hydrogen peroxide, and peroxide acetic acid against V. parahaemolyticus. Ten strains of V. parahaemolyticus isolated from Korean foods and 4 strains of standard V. parahaemolyticus were compared for efficacies of various sanitizers and disinfectants by EN 1276 method based on quantitative suspension test. Ethanol (75 and 95%), 25 ppm of iodine, 100 ppm of quaternary ammonium, 145 ppm of hydrogen peroxide and acid showed more than $5log_{10}CFU/mL$ reduction in both clean and dirty conditions. Tests result of chlorine (100 ppm) showed more than $5log_{10}CFU/mL$ reduction in clean condition. Iodine (15 ppm) showed more than $5log_{10}CFU/mL$ reduction except 4 isolated and 1 standard V. parahaemolyticus in clean condition. iodine (15 ppm) also showed under $5log_{10}CFU/mL$ reduction $(0.93{\sim}3.73log_{10}CFU/mL)$ in dirty condition. Eleven hundred ppm of hydrogen peroxide was evaluated as weak sanitizer and disinfectant due to their $0.99{\sim}4.79log_{10}CFU/mL$ reduction on both clean and dirty conditions. Consequently, ethanol, iodine (25 ppm), chlorine (200 ppm), quaternary ammonium, acid and peroxide acetic acid were thought to be effective sanitizer and disinfectant against V. parahaemolyticus.

Combination of Hydrophobic Filtration and Enrichment Methods for Detecting Bacillus cereus in Fresh-Cut Cabbage

  • Lee, Sujung;Choi, Yukyung;Lee, Heeyoung;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.33 no.5
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    • pp.325-329
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    • 2018
  • This study developed a rapid detection method for Bacillus cereus in fresh-cut cabbages. Fresh-cut cabbage samples were inoculated at 1-, 2- and 3-Log CFU/g, and pathogens were enriched in tryptic soy broth containing 0.15% polymyxin B at $30^{\circ}C$, $37^{\circ}C$, and $42^{\circ}C$ to determine the detection limit and appropriate enrichment temperature for multiplex PCR detection. Enriched bacterial cells in enrichment broth were collected in a hydrophobic filter prior to DNA extraction for multiplex PCR. Filters were resuspended in distilled water, and DNA was extracted from the suspension. DNA samples were further analyzed by multiplex PCR. Detection limit of multiplex PCR was 5-Log CFU/mL. B. cereus cell counts were higher (P < 0.05) at $42^{\circ}C$ than other temperatures. Detection rate of 1-, 2-, and 3-Log CFU/g inoculated samples were 60%, 80%, and 100% after enrichment respectively. However, when enriched samples were filtered with hydrophobic membrane filter, detection rates became 100%, regardless of inoculation level. Results indicate a combination of enrichment with hydrophobic filtration improves rapid detection efficiency of B. cereus in fresh-cut cabbage by multiplex PCR.

Surface roughness and $Candida$ $albicans$ adhesion to flexible denture base according to various polishing methods (연마방법에 따른 탄성의치의 표면거칠기와 $Candida$ $albicans$의 부착율 변화)

  • Oh, Ju-Won;Seo, Jae-Min;Ahn, Seung-Keun;Park, Ju-Mi;Kang, Cheol-Kyun;Song, Kwang-Yeob
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.2
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    • pp.106-111
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    • 2012
  • Purpose: The purpose of this study was to compare the effect of 3 chairside polishing methods and laboratory polishing methods on surface roughness and $C.$ $albicans$ adhesion of polyamide denture base. Materials and methods: Using contact profilometer, the surface of polyamide specimens ($25{\times}15{\times}2mm$) was studied after conventional polishing without finishing and after chiarside polishing with 2 chiarside polishing kits and chairside-pumice polishing following finishing with tungsten carbide bur. To evaluate the adhesion of $C.$ $albicans$, $C.$ $albicans$ suspension was overlayed on the test specimen. And the specimens were incubated for 2 hours. Imprint culture method was achieved and counted the colony on the agar plate. Polished polyamide were evaluated using a scanning electron microscope. The statistics were conducted using one-way ANOVA and in case of difference, Scheffe test and Tamhane's T2 test were used. Results: Surface roughness (Ra) of surfaces polished with 2 chairside polishing kits had higher than conventional polishing and pumice polishing. The highest roughness value was $0.32{\pm}0.10{\mu}m$, and the lowest was $0.02{\pm}0.00{\mu}m$. The adhesion of $C.$ $albicans$ on the specimens polished with chairside polishing group and pumice polishing group were increased than conventional polishing group ($P$<.01). Conclusion: Conventional laboratory polishing was found to produce the smoothest surface and the lowest adhesion of $C.$ $albicans$. Two groups polished with Chairside polishing kits were similar with respect to surface roughness. Surface of the specimen polished with pumice is significantly smoother than 2 chairside polishing groups, but the result of $C.$ $albicans$ adhesion is that group polished with pumice was similar with 2 chairside polishing groups ($P$>.01).

Particulate Matter from Asian Dust Storms Induces the Expression of Proinflammatory Cytokine in A549 Epithelial Cells (PM10이 A549 Cells에서 전염증성 Cytokine발현에 미치는 영향)

  • Kim, Jung Ho;Jeon, Hyo Keun;Kim, Mi Kyeong;Kyung, Sun Yong;An, Chang Hyeok;Lee, Sang Pyo;Park, Jung Woong;Jeong, Sung Hwan
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.6
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    • pp.663-672
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    • 2006
  • Background: $PM_{10}$(Particulate matter with a diameter ($<10{\mu}m$), which is characterized by different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also produce unique particulate matter in affected areas. This study investigated the cytokine produced by A549 epithelial cells exposed to particles collected during both the Asian dust pfenomenon and ambient air particles in a non-dusty period. Method: Air samples were collected using a high volume air sampler(Sibata Model HV500F) with an air flow at $500{\ell}/min$ for at least 6 hours. The cytokine messenger RNA(mRNA) was measured using a reverse transcriptase polymerase chain reaction(RT-PCR). The A549 cells were exposed to 10 to $500{\mu}g/m{\ell}$ of a suspension containing $PM_{10}$ for 24 hours. Each was compared with those in the non-exposed control cells. Result: The mRNA levels of interleukin(IL)-$1{\alpha}$, $IL-I{\beta}$, IL-8, and the granulocyte macrophage colony stimulating factor(GM-CSF) increased after veing exposed to $PM_{10}$ in the ambient air particles, compared with those in the non-exposed control cells. The increase in $IL-1{\alpha}$ and IL-8 were dose dependent at a $PM_{10}$ concentration between $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$. The mRNA level of IL-8 in the A549 epithelial cells was higher during the in the Asian dust period($500{\mu}g/m{\ell}$) than during the non dust period. Conclusion: A549 cells exposed to the $PM_{10}$ collected during the Asian dust period produce more proinflammatory cytokine than during non-dusty period. This cytokine enhances the local inflammatory response in the airways and can also contribute to the systemic component of this inflammatory process.

Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Approaches on Optimum Conditions for Agrobacterium-Mediated Transformation of Phalaenopsis (호접란의 Agrobacterium 이용 형질전환 시스템의 최적조건 구명을 위한 연구)

  • Na, Ae Sil;Been, Chul Gu;Jeong, Byoung Ryong
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.1
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    • pp.1-8
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    • 2010
  • Sensitivities of PLBs of four Phalaenopsis cultivars, P. 'Taisuco Windian', P. 'Nancy Amour', P. 'Pink Twilight' and P. 'Taipei Gold' to kanamycin, spectinomycin and hygromycin at different concentrations (0, 25, 50, 100, 200, and $400mg{\cdot}L^{-1}$) were examined. Hygromycin was favorable for selecting the transformants in the genetic transformation of Phalaenopsis as PLBs of four cultivars were all dead at even $25mg{\cdot}L^{-1}$ hygromycin. Responses of PLBs of P. 'Maki Watanabe' and P. 'Brother Lawrence' to DL-phosphinothricin (PPT) were determined at different concentrations (0, 0.1. 0.25, 0.5, 1.0, 2.0, 2.5, and $5.0mg{\cdot}L^{-1}$) and $0.5mg{\cdot}L^{-1}$ PPT was thought to be suitable for selecting the transformants of Phalaenopsis. The optimum conditions for Agrobacterium cocultivation with Phalaenopsis PLBs were examined using a two-step cocultivation method in Dtps. 'City Girl' and A. tumefaciens LBA4404. In the first infection period in a 1 : 10 suspension of Agrobacterium to a VW medium, 1 hr infection showed the highest PLB survival ratio. And then, PLBs were cocultivated with a bacterial strain and a 3-day cocultivation period was better for Phalaenopsis PLBs than a prolonged period. Agrobacterium tumefaciens strains LBA4404 (pTOK233) and EHA105 (pGA643) were used to compare their efficiency on the genetic transformation of Phalaenopsis PLBs. The PLBs infected with EHA105 survived more than those infected with LBA4404 after two days in a dark condition and two weeks in light condition on a selective medium. About 1,000 PLBs for each of P. 'Maki Watanabe' and P. 'Brother Lawrence', and each bacterial strain of AGL1 (pCAMBIA3301) and LBA4404 (pTOK233) were used for the regeneration of transgenic plants. The bacterial strain AGL1 had a higher genetic transformation efficiency than LBA4404, with no significant difference between cultivars. In this study, 11 hygromycin-resistant plantlets and 32 PPT-resistant plantlets were produced, but these putative transgenic plantlets need further examinations.