| 1 | Identification of stable normalization genes for quantitative real-time PCR in porcine articular cartilage显示文摘Background: Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes. Results: Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage. The genes evaluated were: beta actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hydroxymethylbilane synthase, hypoxanthine phosphoribosyl transferase, peptidylprolyl isomerase A (cyclophilin A), ribosomal protein L4, succinate dehydrogenase flavoprotein subunit A, TATA box binding protein, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein-zeta polypeptide. The stability of the genes was measured using geNorm, BestKeeper, and NormFinder software. The four most stable genes measured via geNorm were (most to least stable) succinate dehydrogenase flavoprotein, subunit A, peptidylprolyl isomerase A, glyceraldehyde-3-phosphate dehydrogenase, beta actin; the four most stable genes measured via BestKeeper were glyceraldehyde-3-phosphate dehydrogenase, peptidylprolyl isomerase A, beta actin, succinate dehydrogenase flavoprotein, subunit A; and the four most stable genes measured via NormFinder were peptidylprolyl isomerase A, succinate dehydrogenase flavoprotein, subunit A, glyceraldehyde-3-phosphate dehydrogenase, beta actin. Conclusions: BestKeeper, geNorm, and NormFinder all generated similar results for the most stable genes in porcine articular cartilage. The use of these appropriate reference genes will facilitate accurate gene expression studies of porcine articular cartilage and suggest appropriate housekeeping genes for articular cartilage studies in other species. | Ryan S McCulloch Melissa S Ashwell Audrey T O'Nan Peter L Mente | 2012 | Journal of Animal Science and Biotechnology2012,3,4: | 2 |
| 3 | Fecal microbial diversity and putative function in captive western lowland gorillas(Gorilla gorilla gorilla),common chimpanzees(Pan troglodytes),Hamadryas baboons(Papio hamadryas)and binturongs(Arctictis binturong)显示文摘Microbial populations in the gastrointestinal tract contribute to host health and nutrition.Although gut microbial ecology is well studied in livestock and domestic animals,little is known of the endogenous populations inhabiting primates or carnivora.We characterized microbial populations in fecal cultures from gorillas(Gorilla gorilla gorilla),common chimpanzees(Pan troglodytes),Hamadryas baboons(Papio hamadryas)and binturongs(Arctictis binturong)to compare the microbiomes associated with different gastrointestinal morphologies and different omnivorous feeding strategies.Each species was fed a distinct standardized diet for 2 weeks prior to fecal collection.All diets were formulated to reflect the species’feeding strategies in situ.Fresh fecal samples were pooled within species and used to inoculate in vitro batch cultures.Acetate,propionate,butyrate and valerate were measured after 24 h of incubation.Eubacterial DNA was extracted from individual fecal samples,pooled,and the cpn60 gene region was amplified and then sequenced to identify the major eubacterial constituents associated with each host species.Short chain fatty acids(P<0.001)and methane(P<0.001)were significantly different across species.Eubacterial profiles were consistent with fermentation data and suggest an increase in diversity with dietary fiber. | Erin A.McKENNEY Melissa ASHWELL Joanna E.LAMBERT Vivek FELLNER | 2014 | Integrative Zoology2014,9,5: | 1 |