• Title/Summary/Keyword: equine

Search Result 334, Processing Time 0.024 seconds

GAPDH, β-actin and β2-microglobulin, as three common reference genes, are not reliable for gene expression studies in equine adipose- and marrow-derived mesenchymal stem cells

  • Nazari, Fatemeh;Parham, Abbas;Maleki, Adham Fani
    • Journal of Animal Science and Technology
    • /
    • v.57 no.5
    • /
    • pp.18.1-18.8
    • /
    • 2015

  • Background: Quantitative real time reverse transcription PCR (qRT-PCR) is one of the most important techniques for gene-expression analysis in molecular based studies. Selecting a proper internal control gene for normalizing data is a crucial step in gene expression analysis via this method. The expression levels of reference genes should be remained constant among cells in different tissues. However, it seems that the location of cells in different tissues might influence their expression. The purpose of this study was to determine whether the source of mesenchymal stem cells (MSCs) has any effect on expression level of three common reference genes (GAPDH, ${\beta}$-actin and ${\beta}2$-microglobulin) in equine marrow- and adipose-derived undifferentiated MSCs and consequently their reliability for comparative qRT-PCR. Materials and methods: Adipose tissue (AT) and bone marrow (BM) samples were harvested from 3 mares. MSCs were isolated and cultured until passage 3 (P3). Total RNA of P3 cells was extracted for cDNA synthesis. The generated cDNAs were analyzed by quantitative real-time PCR. The PCR reactions were ended with a melting curve analysis to verify the specificity of amplicon. Results: The expression levels of GAPDH were significantly different between AT- and BM-derived MSCs (p < 0.05). Differences in expression level of ${\beta}$-actin (P < 0.001) and B2M (P < 0.006.) between MSCs derived from AT and BM were substantially higher than GAPDH. In addition, the fold change in expression levels of GAPDH, ${\beta}$-actin and B2M in AT-derived MSCs compared to BM-derived MSCs were 2.38, 6.76 and 7.76, respectively. Conclusion: This study demonstrated that GAPDH and especially ${\beta}$-actin and B2M express in different levels in equine AT- and BM-derived MSCs. Thus they cannot be considered as reliable reference genes for comparative quantitative gene expression analysis in MSCs derived from equine bone marrow and adipose tissue.

  • https://doi.org/10.1186/s40781-015-0050-8 인용 KSCI

Biological Activities of Tethered Equine Chorionic Gonadotropin (eCG) and Its Deglycosylated Mutants

  • ;;;;N.P JarGil
    • Proceedings of the KSAR Conference
    • /
    • 2004.06a
    • /
    • pp.221-221
    • /
    • 2004

  • Equine chorionic gonadotropin (eCG), which consists of highly glycosylated α- and β-subunits, is a unique member of the gonadotropin family because it elicits response characteristics of both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in other species than the horse. To determine whether α and β subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule was constructed and transfected into Chinese hamster ovary (CHO-K1) cells. (omitted)

  • PDF

The Effects of Solvents in Reservior Solution on Protein Crystallization Using Vapor Diffusion method (증기확산법에 의한 단백질 결정화에 미치는 Reservicr 용액의 영향)

  • 이정희;정용제
    • Korean Journal of Crystallography
    • /
    • v.5 no.1
    • /
    • pp.33-38
    • /
    • 1994

  • 'Heterogeneous' vapor diffusion experiments were carried out using hen egg white Iysozyme and equine serum albumin as model proteins: droplets were equilibrated against reservoir solutiorls containing an alternative precipitant which is different from results showed that the use of polyethylene glycol as an alternative precipitant instead of NaCl or ammonium sulfate reduces equilibration rate between droplet and reservoir solution. By using the heterogeneous vapor diffusion techniqlue it is possible to control the equilibration rate by adjusting the relative amounts of ionic salts and nonionic yecipitants in reservoir solutions.

  • PDF