Selective measurement of α smooth muscle actin: why β-actin can not be used as a housekeeping gene when tissue fibrosis occurs.


BACKGROUND:Prevalence of fibroproliferative diseases, including chronic kidney disease is rapidly increasing and has become a major public health problem worldwide. Fibroproliferative diseases are characterized by increased expression of α smooth muscle actin (α-SMA) that belongs to the family of the six conserved actin isoforms showing high degree homology. The aim of the present study was to develop real-time PCRs that clearly discriminate α-SMA and ß-actin from other actin isoforms. RESULTS:Real-time PCRs using self-designed mouse, human and rat specific α-SMA or ß-actin primer pairs resulted in the specific amplification of the artificial DNA templates corresponding to mouse, human or rat α-SMA or ß-actin, however ß-actin showed cross-reaction with the housekeeping γ-cyto-actin. We have shown that the use of improperly designed literary primer pairs significantly affects the results of PCRs measuring mRNA expression of α-SMA or ß-actin in the kidney of mice underwent UUO. CONCLUSION:We developed a set of carefully designed primer pairs and PCR conditions to selectively determine the expression of mouse, human or rat α-SMA and ß-actin isoforms. We demonstrated the importance of primer specificity in experiments where the results are normalized to the expression of ß-actin especially when fibrosis and thus increased expression of α-SMA is occur.


BMC Mol Biol


BMC molecular biology


Veres-Székely A,Pap D,Sziksz E,Jávorszky E,Rokonay R,Lippai R,Tory K,Fekete A,Tulassay T,Szabó AJ,Vannay Á




Has Abstract


2017-04-27 00:00:00












  • Reference loci for RT-qPCR analysis of differentiating human embryonic stem cells.

    abstract:BACKGROUND:Selecting stably expressed reference genes is essential for proper reverse transcription quantitative polymerase chain reaction gene expression analysis. However, this choice is not always straightforward. In the case of differentiating human embryonic stem (hES) cells, differentiation itself introduces chan...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Vossaert L,O'Leary T,Van Neste C,Heindryckx B,Vandesompele J,De Sutter P,Deforce D

    更新日期:2013-09-12 00:00:00

  • SPRR2A enhances p53 deacetylation through HDAC1 and down regulates p21 promoter activity.

    abstract:BACKGROUND:Small proline rich protein (SPRR) 2A is one of 14 SPRR genes that encodes for a skin cross-linking protein, which confers structural integrity to the cornified keratinocyte cell envelope. New evidence, however, shows that SPRR2A is also a critical stress and wound repair modulator: it enables a variety of ba...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Mizuguchi Y,Specht S,Lunz JG 3rd,Isse K,Corbitt N,Takizawa T,Demetris AJ

    更新日期:2012-06-25 00:00:00

  • Phylogeny and expression of carbonic anhydrase-related proteins.

    abstract:BACKGROUND:Carbonic anhydrases (CAs) are found in many organisms, in which they contribute to several important biological processes. The vertebrate alpha-CA family consists of 16 subfamilies, three of which (VIII, X and XI) consist of acatalytic proteins. These are named carbonic anhydrase related proteins (CARPs), an...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Aspatwar A,Tolvanen ME,Parkkila S

    更新日期:2010-03-31 00:00:00

  • Transcription of the rat testis-specific Rtdpoz-T1 and -T2 retrogenes during embryo development: co-transcription and frequent exonisation of transposable element sequences.

    abstract:BACKGROUND:Retrotransposition is an important evolutionary force for the creation of new and potentially functional intronless genes which are collectively called retrogenes. Many retrogenes are expressed in the testis and the gene products have been shown to actively participate in spermatogenesis and other unique fun...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Huang CJ,Lin WY,Chang CM,Choo KB

    更新日期:2009-07-25 00:00:00

  • Small nucleolar RNA Sf-15 regulates proliferation and apoptosis of Spodoptera frugiperda Sf9 cells.

    abstract:BACKGROUND:Small nucleolar RNAs (snoRNAs) function in guiding 2'-O-methylation and pseudouridylation of ribosomal RNAs (rRNAs) and small nuclear RNAs (snRNAs). In recent years, more and more snoRNAs have been found to play novel roles in mRNA regulation, such as pre-mRNA splicing or RNA editing. In our previous study, ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Wu B,Huang L,Qiu W,Liu X,Shen Y,Lu Y,Yang Z,Li X,Cui B,Xu S,Qiao H,Qiu R,Yao L,Kan Y,Li D

    更新日期:2019-04-11 00:00:00

  • Biochemical characterization of a recombinant Japanese encephalitis virus RNA-dependent RNA polymerase.

    abstract:BACKGROUND:Japanese encephalitis virus (JEV) NS5 is a viral nonstructural protein that carries both methyltransferase and RNA-dependent RNA polymerase (RdRp) domains. It is a key component of the viral RNA replicase complex that presumably includes other viral nonstructural and cellular proteins. The biochemical proper...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Kim YG,Yoo JS,Kim JH,Kim CM,Oh JW

    更新日期:2007-07-11 00:00:00

  • Transcriptional response of pancreatic beta cells to metabolic stimulation: large scale identification of immediate-early and secondary response genes.

    abstract:BACKGROUND:Physiological long term adaptation of pancreatic beta cells is driven by stimuli such as glucose and incretin hormones acting via cAMP (e.g. GLP-1) and involves regulated gene expression. Several rapidly inducible immediate-early genes (IEGs) have been identified in beta cells. Many of these IEGs code for tr...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Glauser DA,Brun T,Gauthier BR,Schlegel W

    更新日期:2007-06-22 00:00:00

  • Identification, characterization and expression of novel Sex Hormone Binding Globulin alternative first exons in the human prostate.

    abstract:BACKGROUND:The human Sex Hormone Binding Globulin (SHBG) gene, located at 17p13.1, comprises, at least, two different transcription units regulated by two different promoters. The first transcription unit begins with the exon 1 sequence and is responsible for the production of plasma SHBG by the hepatocytes, while the ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Pinós T,Barbosa-Desongles A,Hurtado A,Santamaria-Martínez A,de Torres I,Morote J,Reventós J,Munell F

    更新日期:2009-06-17 00:00:00

  • Preliminary molecular characterization of the human pathogen Angiostrongylus cantonensis.

    abstract:BACKGROUND:Human angiostrongyliasis is an emerging food-borne public health problem, with the number of cases increasing worldwide, especially in mainland China. Angiostrongylus cantonensis is the causative agent of this severe disease. However, little is known about the genetics and basic biology of A. cantonensis. R...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: He H,Cheng M,Yang X,Meng J,He A,Zheng X,Li Z,Guo P,Pan Z,Zhan X

    更新日期:2009-10-25 00:00:00

  • Structural and functional analysis of four non-coding Y RNAs from Chinese hamster cells: identification, molecular dynamics simulations and DNA replication initiation assays.

    abstract:BACKGROUND:The genes coding for Y RNAs are evolutionarily conserved in vertebrates. These non-coding RNAs are essential for the initiation of chromosomal DNA replication in vertebrate cells. However thus far, no information is available about Y RNAs in Chinese hamster cells, which have already been used to detect repli...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: de Lima Neto QA,Duarte Junior FF,Bueno PS,Seixas FA,Kowalski MP,Kheir E,Krude T,Fernandez MA

    更新日期:2016-01-05 00:00:00

  • A modification of Representational Difference Analysis, with application to the cloning of a candidate in the Reelin signalling pathway.

    abstract:BACKGROUND:cDNA-RDA is one of the subtractive cloning techniques used to isolate differentially expressed genes between two complex cDNA populations. In the present study we present a modification of the protocol described by Hubank and Schatz. RESULTS:In the post-hybridization mix, the 5'-ends of homoduplexes of inte...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Kuvbachieva AA,Goffinet AM

    更新日期:2002-04-24 00:00:00

  • Characterization, phylogeny, alternative splicing and expression of Sox30 gene.

    abstract:BACKGROUND:Members of the Sox gene family isolated from both vertebrates and invertebrates have been proved to participate in a wide variety of developmental processes, including sex determination and differentiation. Among these members, Sox30 had been considered to exist only in mammals since its discovery, and its e...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Han F,Wang Z,Wu F,Liu Z,Huang B,Wang D

    更新日期:2010-12-11 00:00:00

  • GATA elements control repression of cardiac troponin I promoter activity in skeletal muscle cells.

    abstract:BACKGROUND:We reported previously that the cardiac troponin I (cTnI) promoter drives cardiac-specific expression of reporter genes in cardiac muscle cells and in transgenic mice, and that disruption of GATA elements inactivates the cTnI promoter in cultured cardiomyocytes. We have now examined the role of cTnI promoter...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Di Lisi R,Picard A,Ausoni S,Schiaffino S

    更新日期:2007-09-17 00:00:00

  • Translation initiation region sequence preferences in Escherichia coli.

    abstract:BACKGROUND:The mRNA translation initiation region (TIR) comprises the initiator codon, Shine-Dalgarno (SD) sequence and translational enhancers. Probably the most abundant class of enhancers contains A/U-rich sequences. We have tested the influence of SD sequence length and the presence of enhancers on the efficiency o...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Vimberg V,Tats A,Remm M,Tenson T

    更新日期:2007-10-31 00:00:00

  • Identification of ColR binding consensus and prediction of regulon of ColRS two-component system.

    abstract:BACKGROUND:Conserved two-component system ColRS of Pseudomonas genus has been implicated in several unrelated phenotypes. For instance, deficiency of P. putida ColRS system results in lowered phenol tolerance, hindrance of transposition of Tn4652 and lysis of a subpopulation of glucose-grown bacteria. In order to disco...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Kivistik PA,Kivi R,Kivisaar M,Hõrak R

    更新日期:2009-05-16 00:00:00

  • Parental origin of chromosomes influences crossover activity within the Kcnq1 transcriptionally imprinted domain of Mus musculus.

    abstract:BACKGROUND:Among the three functions of DNA, mammalian replication and transcription can be subject to epigenetic imprinting specified by the parental origin of chromosomes, and although there is suggestive indication that this is also true for meiotic recombination, no definitive evidence has yet been reported. RESUL...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Ng SH,Madeira R,Parvanov ED,Petros LM,Petkov PM,Paigen K

    更新日期:2009-05-13 00:00:00

  • Ataxia telangiectasia mutated (ATM) interacts with p400 ATPase for an efficient DNA damage response.

    abstract:BACKGROUND:Ataxia telangiectasia mutated (ATM) and TRRAP proteins belong to the phosphatidylinositol 3-kinase-related kinase family and are involved in DNA damage repair and chromatin remodeling. ATM is a checkpoint kinase that is recruited to sites of DNA double-strand breaks where it phosphorylates a diverse range of...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Smith RJ,Savoian MS,Weber LE,Park JH

    更新日期:2016-11-04 00:00:00

  • The yfhQ gene of Escherichia coli encodes a tRNA:Cm32/Um32 methyltransferase.

    abstract:BACKGROUND:Naturally occurring tRNAs contain numerous modified nucleosides. They are formed by enzymatic modification of the primary transcripts during the complex RNA maturation process. In model organisms Escherichia coli and Saccharomyces cerevisiae most enzymes involved in this process have been identified. Interes...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Purta E,van Vliet F,Tkaczuk KL,Dunin-Horkawicz S,Mori H,Droogmans L,Bujnicki JM

    更新日期:2006-07-18 00:00:00

  • USF2 inhibits C/EBP-mediated transcriptional regulation of the RIIbeta subunit of cAMP-dependent protein kinase.

    abstract:BACKGROUND:Cyclic AMP-dependent protein kinase (PKA) plays a central role in regulation of energy metabolism. Upon stimulation of testicular Sertoli cells by follicle stimulating hormone (FSH), glycolysis is activated to increase the production of nutrients for the germ cells, and a new regulatory subunit of cAMP-depen...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Dahle MK,Taskén K,Taskén KA

    更新日期:2002-06-21 00:00:00

  • Deficiency of Pten accelerates mammary oncogenesis in MMTV-Wnt-1 transgenic mice.

    abstract:BACKGROUND:Germline mutations in the tumor suppressor PTEN predispose human beings to breast cancer, and genetic and epigenetic alterations of PTEN are also detected in sporadic human breast cancer. Germline Pten mutations in mice lead to the development of a variety of tumors, but mammary carcinomas are infrequently f...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Li Y,Podsypanina K,Liu X,Crane A,Tan LK,Parsons R,Varmus HE

    更新日期:2001-01-01 00:00:00

  • Alternative splicing and differential subcellular localization of the rat FGF antisense gene product.

    abstract:BACKGROUND:GFG/NUDT is a nudix hydrolase originally identified as the product of the fibroblast growth factor-2 antisense (FGF-AS) gene. While the FGF-AS RNA has been implicated as an antisense regulator of FGF-2 expression, the expression and function of the encoded GFG protein is largely unknown. Alternative splicing...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Zhang SC,MacDonald KA,Baguma-Nibasheka M,Geldenhuys L,Casson AG,Murphy PR

    更新日期:2008-01-23 00:00:00

  • RE1 silencing transcription factor (REST) negatively regulates ALL1-fused from chromosome 1q (AF1q) gene transcription.

    abstract:BACKGROUND:ALL1-fused from chromosome 1q (AF1q), originally considered as an oncogenic factor, has been implicated in neuronal development; however, its upstream regulatory mechanisms in neural system remained elusive. RESULTS:Our study showed that REST (RE1 silencing transcription factor), a key transcription factor ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Hu Y,Sun Q,Zhang C,Sha Q,Sun X

    更新日期:2015-09-05 00:00:00

  • A novel, non-radioactive eukaryotic in vitro transcription assay for sensitive quantification of RNA polymerase II activity.

    abstract:BACKGROUND:Many studies of the eukaryotic transcription mechanism and its regulation rely on in vitro assays. Conventional RNA polymerase II transcription assays are based on radioactive labelling of the newly synthesized RNA. Due to the inefficient in vitro transcription, the detection of the RNA involving purificatio...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Voss C,Schmitt B,Werner-Simon S,Lutz C,Simon W,Anderl J

    更新日期:2014-04-03 00:00:00

  • Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces cerevisiae.

    abstract:BACKGROUND:Real-time RT-PCR is the recommended method for quantitative gene expression analysis. A compulsory step is the selection of good reference genes for normalization. A few genes often referred to as HouseKeeping Genes (HSK), such as ACT1, RDN18 or PDA1 are among the most commonly used, as their expression is a...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Teste MA,Duquenne M,François JM,Parrou JL

    更新日期:2009-10-30 00:00:00

  • Analysis of the regulation of fatty acid binding protein 7 expression in human renal carcinoma cell lines.

    abstract:BACKGROUND:Improving the treatment of renal cell carcinoma (RCC) will depend on the development of better biomarkers for predicting disease progression and aiding the design of appropriate therapies. One such marker may be fatty acid binding protein 7 (FABP7), also known as B-FABP and BLBP, which is expressed normally ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Takaoka N,Takayama T,Teratani T,Sugiyama T,Mugiya S,Ozono S

    更新日期:2011-07-19 00:00:00

  • RNF168, a new RING finger, MIU-containing protein that modifies chromatin by ubiquitination of histones H2A and H2AX.

    abstract:BACKGROUND:Modulation of chromatin structure has emerged as a critical molecular device to control gene expression. Histones undergo different post-translational modifications that increase chromatin accessibility to a number of regulatory factors. Among them, histone ubiquitination appears relevant in nuclear processe...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Pinato S,Scandiuzzi C,Arnaudo N,Citterio E,Gaudino G,Penengo L

    更新日期:2009-06-05 00:00:00

  • Evaluation of reference genes for real-time quantitative PCR studies in Candida glabrata following azole treatment.

    abstract:BACKGROUND:The selection of stable and suitable reference genes for real-time quantitative PCR (RT-qPCR) is a crucial prerequisite for reliable gene expression analysis under different experimental conditions. The present study aimed to identify reference genes as internal controls for gene expression studies by RT-qPC...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Li QQ,Skinner J,Bennett JE

    更新日期:2012-06-29 00:00:00

  • TRPC1 transcript variants, inefficient nonsense-mediated decay and low up-frameshift-1 in vascular smooth muscle cells.

    abstract:BACKGROUND:Transient Receptor Potential Canonical 1 (TRPC1) is a widely-expressed mammalian cationic channel with functional effects that include stimulation of cardiovascular remodelling. The initial aim of this study was to investigate variation in TRPC1-encoding gene transcripts. RESULTS:Extensive TRPC1 transcript ...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Dedman AM,Majeed Y,Tumova S,Zeng F,Kumar B,Munsch C,Bateson AN,Wittmann J,Jäck HM,Porter KE,Beech DJ

    更新日期:2011-07-12 00:00:00

  • Genome-wide identification of brain miRNAs in response to high-intensity intermittent swimming training in Rattus norvegicus by deep sequencing.

    abstract:BACKGROUND:Physical exercise can improve brain function by altering brain gene expression. The expression mechanisms underlying the brain's response to exercise still remain unknown. miRNAs as vital regulators of gene expression may be involved in regulation of brain genes in response to exercise. However, as yet, very...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Zhao Y,Zhang A,Wang Y,Hu S,Zhang R,Qian S

    更新日期:2019-01-15 00:00:00

  • Comparative in silico analysis identifies bona fide MyoD binding sites within the Myocyte stress 1 gene promoter.

    abstract:BACKGROUND:Myocyte stress 1 (MS1) is a striated muscle actin binding protein required for the muscle specific activity of the evolutionary ancient myocardin related transcription factor (MRTF)/serum response factor (SRF) transcriptional pathway. To date, little is known about the molecular mechanisms that govern skelet...

    journal_title:BMC molecular biology

    pub_type: 杂志文章


    authors: Ounzain S,Dacwag CS,Samani NJ,Imbalzano AN,Chong NW

    更新日期:2008-05-19 00:00:00