A validation of 10 feline reference genes for gene expression measurements in snap-frozen tissues

Penning, Louis C.; Vrieling, H.E.; Brinkhof, Bas; Riemers, Frank M.; Rothuizen, J.; Rutteman, Gerard R.; Hazewinkel, H. A. W.

doi:10.1016/j.vetimm.2007.08.006

Cited by 64 publications

(79 citation statements)

References 44 publications

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“…Moreover, only one reference gene for real-time RT-PCR normalization was used in most of the studies of gene transcription in fish. The selection of two reference genes, however, is prevalent in the non-fish literature (Li et al 2005;Nygard et al 2007;Penning et al 2007). …”

Section: Discussionmentioning

confidence: 99%

Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

Guo

Gao

et al. 2014

Fish Physiol Biochem

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Real-time reverse transcriptase (RT) polymerase chain reaction (PCR) requires data normalization using an appropriate reference gene in order to obtain more reliable results with biological significance. We cloned a partial sequence of elongation factor-1-α (EF1α) and ribosomal protein L17 (RPL17) from Monopterus albus. We investigated the suitability of five commonly used reference genes [18S ribosomal RNA (18S), cytoskeletal protein (β-actin), glyceraldehyde phosphate dehydrogenase (GAPDH), EF1α and RPL17] as potential quantitative reference genes for normalizing real-time RT-PCR data generated in gonads of different developmental stages and in other tissues of M. albus. Analysis of the data indicated that 18S, β-actin and GAPDH are not suitable as reference genes because of their levels of variations of expression. EF1α and RPL17 might be suitable as reference genes in the gonads of different developmental stages as well as in other tissues of M. albus.

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Section: Discussionmentioning

confidence: 99%

Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

Guo

Gao

et al. 2014

Fish Physiol Biochem

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“…This selection was based on reference genes already described and evaluated in companion animals such as dogs and cats (Brinkhof et al, 2006;Penning et al, 2007), horse skin (Bogaert et al, 2006), dolphin skin (Spinsanti et al, 2006) and human skin samples (de Kok et al, 2005). This low number of papers on this specific subject further shows that the evaluation of reference genes for skin specimen is at its infancy.…”

Section: Introductionmentioning

confidence: 99%

A GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitis

Schlotter

Veenhof

Brinkhof

et al. 2009

Veterinary Immunology and Immunopathology

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“…New primers were designed as needed using the Integrated DNA Technologies' primer design tools based on the genome of the domestic cat ( Felis catus ). TLR10 primer sequences were obtained from Mercier et al (2012) and feline primer sequences for the reference gene HPRT were obtained from Penning et al (2007). PCR products for each gene were purified using a PCR clean-up system (Promega, #A9281) and sequenced at the MSU Research Technology Support Facility.…”

Section: Methodsmentioning

confidence: 99%

Characterization of toll-like receptors 1–10 in spotted hyenas

et al. 2014

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Previous research has shown that spotted hyenas (Crocuta crocuta) regularly survive exposure to deadly pathogens such as rabies, canine distemper virus, and anthrax, suggesting that they have robust immune defenses. Toll-like receptors (TLRs) recognize conserved molecular patterns and initiate a wide range of innate and adaptive immune responses. TLR genes are evolutionarily conserved, and assessing TLR expression in various tissues can provide insight into overall immunological organization and function. Studies of the hyena immune system have been minimal thus far due to the logistical and ethical challenges of sampling and preserving the immunological tissues of this and other long-lived, wild species. Tissue samples were opportunistically collected from captive hyenas humanely euthanized for a separate study. We developed primers to amplify partial sequences for TLRs 1-10, sequenced the amplicons, compared sequence identity to those in other mammals, and quantified TLR expression in lymph nodes, spleens, lungs, and pancreases. Results show that hyena TLR DNA and protein sequences are similar to TLRs in other mammals, and that TLRs 1-10 were expressed in all tissues tested. This information will be useful in the development of new assays to understand the interactions among the hyena immune system, pathogens, and the microbial communities that inhabit hyenas.

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A validation of 10 feline reference genes for gene expression measurements in snap-frozen tissues

Cited by 64 publications

References 44 publications

Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

A GeNorm algorithm-based selection of reference genes for quantitative real-time PCR in skin biopsies of healthy dogs and dogs with atopic dermatitis

Characterization of toll-like receptors 1–10 in spotted hyenas

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