• Title/Summary/Keyword: 일시적인 형질전환 분석

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Study on PPARγ-dependent adipogenesis regulation by testosterone using transient transfection assays (일시적인 형질전환 분석을 이용한 testosterone에 의한 PPARγ-의존적 지방세포생성 조절에 관한 연구)

  • Jeong, Sunhyo
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.2
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    • pp.482-487
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    • 2014
  • This study is to investigate the effects of testosterone on adipogenesis and its molecular mechanism using RT-PCR analysis and transient transfection assays. Castrated(CAST) mice treated with testosterone had lower white adipose tissue weights and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) than CAST control mice. Consistent with the in vivo data, testosterone treatment inhibited triglyceride accumulation and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) in differentiated 3T3-L1 cells compared with control group. Testosterone-activated androgen receptor(AR) repressed the luciferase reporter gene activity induced by $PPAR{\gamma}$ transfection. Thus, these results suggest that testosterone downregulates the actions of $PPAR{\gamma}$ on adipogenesis through AR.

Agrobacterium- mediated Genetic Transformation and Plant Regeneration of Sweetpotato (Ipomoea batatas) (Agrobacterium 매개에 의한 고구마 형질전환 및 식물체 재분화)

  • Lim, Soon;Yang, Kyoung-Sil;Kwon, Suk-Yoon;Paek, Kee-Yoeup;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.267-271
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    • 2004
  • Transformed sweetpotato (Ipomoea batatas (L.) Lam. cv. Yulmi) plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105/pCAMBIA2301 harboring genes for intron $\beta$-glucuronidase (GUS) and kanamycin resistance. Transient expression of GUS gene was found to be higher when embryogenic calli were co-cultivated with Agrobacterium for 2 days. The co-cultured embryogenic calli transferred to selective MS medium containing 1mg/L 2,4-D, 100mg/L kanamycin, and 400mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 4 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the GUS gene was inserted into the genome of the sweetpotato plants. A histochemical assay revealed that the GUS gene was preferentially expressed in the leaf, petiole, and vascular tissue and tip of root.

Transient Expression of β-gulucuronidase (GUS) gene in Immature Ovules and Calli Derived from Cottonwood Species (Populus deltoides) by Microprojectile Bombardment (포플러의 미성숙(未成熟) 배(胚)와 캘러스에서 유전자총(遺傳子銃)에 의(依)한 GUS-gene의 일시적(一時的) 발현(發現))

  • Kang, Hoduck;Kang, Sang-Gu;Bae, Hanhong;Park, Kyo-Soo;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.86 no.3
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    • pp.261-269
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    • 1997
  • Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and ${\beta}$-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced about $25.75{\pm}2.77$(per ovule), $11.43{\pm}1.22$(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.

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