Document Detail


Efficient extraction of RNA and analysis of gene expression in a long-term Taxus cell culture using real-time RT-PCR.
MedLine Citation:
PMID:  19323277     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A simple, quick and efficient method for isolating total RNA from heavy browning cells was developed by adding polyvinylpyrrolidone, mercaptoethanol and 3 M NaAc during the process of the Trizol (a kind of a widely used RNA extraction buffer) method. High-quality total RNA was isolated and synthesized to cDNA. Transcript levels of four paclitaxel biosynthetic pathway genes: dxr, hmgr, ggpps and dbat were assayed by real-time RT-PCR. The results demonstrated that the transcript levels of these genes experienced a coincident descent in the past three years as well as a decreasing paclitaxel productivity. According to these results, the possible reason for the descending paclitaxel productivity during long-term Taxus media cv. Hicksii cell culture maybe due to a decreasing transcripts level of mass genes in close with a gross secondary metabolite level. Gene manipulation emphasized only on key enzyme genes in the paclitaxel biosynthesis pathway may not hamper the somaclonal variation trend of Taxus media cv. Hicksii cell culture.
Authors:
Li-Qin Li; Chun-Hua Fu; Chun-Fang Zhao; Juan Xia; Wen-Juan Wu; Long-Jiang Yu
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Zeitschrift für Naturforschung. C, Journal of biosciences     Volume:  64     ISSN:  0939-5075     ISO Abbreviation:  Z. Naturforsch., C, J. Biosci.     Publication Date:    2009 Jan-Feb
Date Detail:
Created Date:  2009-03-27     Completed Date:  2009-04-27     Revised Date:  2009-11-04    
Medline Journal Info:
Nlm Unique ID:  8912155     Medline TA:  Z Naturforsch C     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  125-30     Citation Subset:  IM    
Affiliation:
Department of Biology Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, Hubei, China.
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MeSH Terms
Descriptor/Qualifier:
Cell Culture Techniques
DNA Primers
DNA, Bacterial / genetics
Gene Expression Profiling / methods*
Gene Expression Regulation, Plant*
RNA, Plant / genetics*,  isolation & purification
Reverse Transcriptase Polymerase Chain Reaction / methods*
Taxus / cytology,  genetics*
Chemical
Reg. No./Substance:
0/DNA Primers; 0/DNA, Bacterial; 0/RNA, Plant

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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