title: Characterization of reference genes for quantitative real-time PCR analysis in various tissues of Salvia miltiorrhiza.
abstract: Five reference genes, 18S, EF1alpha, alpha-Tubulin, Ubiquitin and Actin, from Salvia miltiorrhiza were analyzed as internal controls for gene expression profiling assay using quantitative real-time polymerase chain reaction (qRT-PCR). The five candidate genes were measured for their transcriptional level in seven tissues, including roots, stems, leaves, sepals, petals, stamens and pistils. Then they were ranked by the GeNorm tool. The results showed that Actin and Ubiquitin were the most stable whereas EF1alpha and 18S did not favor normalization of qRT-PCR results in these tissues. Expression levels of the SmDXR gene were studied in parallel, with Actin and Ubiquitin both or each as reference in the seven tissues, and varying relative quantifications of the SmDXR gene in seven tissues. This study indicated that selection of the most stable genes plays an important role in gene expression profiling assays.
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical College, Beijing, China.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Yang
authors.forename: Shuang
authors.initials: S
authors.lastname: Hou
authors.forename: Guanghong
authors.initials: G
authors.lastname: Cui
authors.forename: Shilin
authors.initials: S
authors.lastname: Chen
authors.forename: Jianhe
authors.initials: J
authors.lastname: Wei
authors.forename: Luqi
authors.initials: L
authors.lastname: Huang
mesh_terms: D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D027561:Salvia miltiorrhiza / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012984:Software; D014158:Transcription, Genetic
chemical_list: D010940:Plant Proteins
references.citation: J Mol Endocrinol. 2000 Oct;25(2):169-93
references.pmid: 11013345
references.citation: Genome Biol. 2002 Jun 18;3(7):RESEARCH0034
references.pmid: 12184808
references.citation: Biotechnol Appl Biochem. 2009 Jan;52(Pt 1):89-95
references.pmid: 18302535
references.citation: BMC Plant Biol. 2004 Aug 18;4:14
references.pmid: 15317655
references.citation: Mol Biol Rep. 2009 Jan;36(1):71-81
references.pmid: 17922220
references.citation: Trends Plant Sci. 2008 Dec;13(12):619-23
references.pmid: 18948055
references.citation: Genome Res. 2002 Feb;12(2):292-7
references.pmid: 11827948
references.citation: Biotechnol Lett. 2003 Nov;25(21):1869-72
references.pmid: 14677714
references.citation: J Virol Methods. 2003 Mar;108(1):1-8
references.pmid: 12565148
references.citation: J Biochem Biophys Methods. 2000 Nov 20;46(1-2):69-81
references.pmid: 11086195
references.citation: Biochem Biophys Res Commun. 2004 Jan 23;313(4):856-62
references.pmid: 14706621
references.citation: J Mol Endocrinol. 2002 Aug;29(1):23-39
references.pmid: 12200227
references.citation: Biochem Biophys Res Commun. 2006 Jun 30;345(2):646-51
references.pmid: 16690022
references.citation: Nucleic Acids Res. 1993 Aug 11;21(16):3809-19
references.pmid: 8367299
references.citation: J Med Microbiol. 2003 May;52(Pt 5):403-408
references.pmid: 12721316
references.citation: J Plant Physiol. 2008 Feb;165(2):203-13
references.pmid: 17257708
references.citation: Biotechniques. 2003 Oct;35(4):740-2, 744, 746 passim
references.pmid: 14579739
references.citation: Trends Plant Sci. 2004 Jan;9(1):49-56
references.pmid: 14729219
references.citation: Comp Biochem Physiol B Biochem Mol Biol. 2001 Oct;130(3):281-9
references.pmid: 11567890
references.citation: J Exp Bot. 2007;58(7):1761-70
references.pmid: 17355947
references.citation: BMC Plant Biol. 2008 Dec 22;8:131
references.pmid: 19102748
references.citation: J Exp Bot. 2005 Nov;56(421):2907-14
references.pmid: 16188960
references.citation: Pharmacol Ther. 2008 Feb;117(2):280-95
references.pmid: 18048101
references.citation: J Clin Virol. 2002 Dec;25 Suppl 3:S47-53
references.pmid: 12467777
references.citation: BMC Mol Biol. 2008 Jun 23;9:59
references.pmid: 18573215
references.citation: DNA Seq. 2005 Apr;16(2):111-20
references.pmid: 16147862
references.citation: Physiol Genomics. 2004 Jul 08;18(2):226-31
references.pmid: 15161965
references.citation: Plant J. 2001 Aug;27(3):213-22
references.pmid: 11532167
references.citation: Planta Med. 2006 Feb;72(3):234-40
references.pmid: 16534728
references.citation: Biotechniques. 2000 Aug;29(2):332-7
references.pmid: 10948434
references.citation: Physiol Genomics. 2000 Apr 27;2(3):143-7
references.pmid: 11015593
references.citation: Plant Cell. 2007 Nov;19(11):3669-91
references.pmid: 18024569
references.citation: J Biotechnol. 1999 Oct 8;75(2-3):291-5
references.pmid: 10617337
references.citation: Plant Physiol. 2002 Aug;129(4):1581-91
references.pmid: 12177470
references.citation: Plant Physiol. 2005 Sep;139(1):5-17
references.pmid: 16166256