• 제목/요약/키워드: DAI

검색결과 2,066건 처리시간 0.03초

Effects of Cell Cycle Regulators on the Cell Cycle Synchronization of Porcine induced Pluripotent Stem Cells

  • Kwon, Dae-Jin;Hwang, In-Sul;Kwak, Tae-Uk;Yang, Hyeon;Park, Mi-Ryung;Ock, Sun-A;Oh, Keon Bong;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • 한국발생생물학회지:발생과생식
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    • 제21권1호
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    • pp.47-54
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    • 2017

  • Unlike mouse results, cloning efficiency of nuclear transfer from porcine induced pluripotent stem cells (piPSCs) is very low. The present study was performed to investigate the effect of cell cycle inhibitors on the cell cycle synchronization of piPSCs. piPSCs were generated using combination of six human transcriptional factors under stem cell culture condition. To examine the efficiency of cell cycle synchronization, piPSCs were cultured on a matrigel coated plate with stem cell media and they were treated with staurosporine (STA, 20 nM), daidzein (DAI, $100{\mu}M$), roscovitine (ROSC, $10{\mu}M$), or olomoucine (OLO, $200{\mu}M$) for 12 h. Flow Cytometry (FACs) data showed that piPSCs in control were in G1 ($37.5{\pm}0.2%$), S ($34.0{\pm}0.6%$) and G2/M ($28.5{\pm}0.4%$). The proportion of cells at G1 in DAI group was significantly higher than that in control, while STA, ROSC and OLO treatments could not block the cell cycle of piPSCs. Both of viability and apoptosis were affected by STA and ROSC treatment, but there were no significantly differences between control and DAI groups. Real-Time qPCR and FACs results revealed that DAI treatment did not affect the expression of pluripotent gene, Oct4. In case of OLO, it did not affect both of viability and apoptosis, but Oct4 expression was significantly decreased. Our results suggest that DAI could be used for synchronizing piPSCs at G1 stage and has any deleterious effect on survival and pluripotency sustaining of piPSCs.

  • https://doi.org/10.12717/DR.2017.21.1.047 인용 PDF KSCI

Agro-infiltration을 이용한 토마토 β-carotene hydroxylase 유전자(ChyB) 과발현 및 담배식물체의 항산화 효과 증진 (Enhanced Antioxident Effect by over Expression of Tomato β-carotene Hydroxylase Gene (ChyB) Using Agrobacterium-infiltration in Tobacco Plant)

  • 최윤정;윤경영;윤해근;서상곤;문용선
    • 원예과학기술지
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    • 제29권3호
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    • pp.267-272
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    • 2011

  • ${\beta}$-carotene hydroxylase는 carotenoids의 생합성 경로에서 astaxanthin 및 zeaxanthin 합성에 관여하는 주요 효소이다. Astaxanthin과 zeaxanthin은 비타민 A 전구체보다 항암효과가 더 높다고 보고되고 있다. 따라서 ${\beta}$-carotene hydroxylase 유전자(ChyB)를 토마토 잎에서 분리하고 binary 벡터에 클로닝 한 후 pIG121-ChyB-tom으로 명명하였다. 토마토 ChyB 유전자를 Agrobacterium-mediated infiltration 방법을 이용하여 생육 8주 된 담배 잎에 일시적 형질전환을 하였다. 감염 0, 1, 2, 3일 후 RT-PCR 한 결과 담배에서는 거의 발현되지 않았던 ${\beta}$-carotene hydroxylase 전사체가 감염 1일 후부터 증가하여 2일 후 최대 전사량을 보이고 3일 후부터 감소하는 경향을 나타내었다. 이 결과를 바탕으로 감염 후 0, 2, 3일째 담배 잎을 채취하여 항산화 효능을 측정한 결과, 감염 2일 후 1,1-diphenyl-pricryl hydrazyl(DPPH) radical 소거 활성이 대조구에 비해 약 30% 증가하는 것을 확인하였다. 이 연구결과를 통해 외래 유용 유전자를 원예작물에 형질전환하여 항산화 및 항암효과가 우수한 zeaxanthin 및 astaxanthin 등 oxy-carotenoids의 함유량을 증진시킨 새로운 기능성 토마토 및 다양한 원예작물의 신품종 개발에 이용될 수 있을 것으로 기대된다.

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