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Journal
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Biochemistry
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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
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Biochemistry
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JoVE Journal
Biochemistry
Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
Please note that all translations are automatically generated.
Click here for the English version.
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
DOI:
10.3791/58166-v
•
09:11 min
•
October 02, 2018
•
Roberto Pech-Kú
,
J. Armando Muñoz-Sánchez
,
Miriam Monforte-González
,
Felipe Vázquez-Flota
,
Beatriz A. Rodas-Junco
,
S.M. Teresa Hernández-Sotomayor
1
Unidad de Bioquímica y Biología Molecular de Plantas
,
Centro de Investigación Científica de Yucatán
,
2
CONACYT, Facultad de Ingeniería Química, Campus de Ciencias Exactas e Ingeniería
,
Universidad Autónoma de Yucatán
Chapters
00:04
Title
00:28
Caffeine Extraction in Cell Suspension of
C. arabica
L.
00:55
Conditions for the Assessment of Caffeine by Thin Layer Chromatography (TLC) Denistometry
01:29
Ribonucleic Acid (RNA) Isolation
02:55
Incubation of RNA Transcript with Deoxyribonuclease (DNase)
03:59
Complementary Deoxyribonucleic Acid (cDNA) Synthesis
05:03
Real-Time Quantitative PCR (qPCR) to Amplify the Gene
CCS1
05:43
Total Protein of Cell Suspensions of
C. arabica
L.
06:26
Enzymatic Assay for Caffeine Synthase
07:39
Results: Enzymatic Activity of Caffeine Synthase Gene Evaluated via qPCR
08:30
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
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。
Tags
Caffeine Extraction
Enzymatic Activity
Gene Expression
Caffeine Synthase
Plant Cell Suspensions
Analytical Chemistry
Biochemistry
Biotechnology
Caffeine Biosynthesis
In Vitro Plant Models
Acetone
TLC
Densitometry
RNA Isolation
Chloroform Isoamyl Alcohol
Phenol
Isopropanol
Ethanol
DEPC-treated Water
DNase
Article
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