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植物细胞悬浮液中咖啡因的提取、酶活性及咖啡因合成酶基因的表达
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
DOI:

09:11 min

October 02, 2018

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Chapters

  • 00:04Title
  • 00:28Caffeine Extraction in Cell Suspension of C. arabica L.
  • 00:55Conditions for the Assessment of Caffeine by Thin Layer Chromatography (TLC) Denistometry
  • 01:29Ribonucleic Acid (RNA) Isolation
  • 02:55Incubation of RNA Transcript with Deoxyribonuclease (DNase)
  • 03:59Complementary Deoxyribonucleic Acid (cDNA) Synthesis
  • 05:03Real-Time Quantitative PCR (qPCR) to Amplify the Gene CCS1
  • 05:43Total Protein of Cell Suspensions of C. arabica L.
  • 06:26Enzymatic Assay for Caffeine Synthase
  • 07:39Results: Enzymatic Activity of Caffeine Synthase Gene Evaluated via qPCR
  • 08:30Conclusion

Summary

自动翻译

本协议描述了一种高效的方法, 用于提取和定量的咖啡因在细胞悬浮液中的表达水平和评价咖啡因合酶酶活性的实验过程。编码这种酶的基因。

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