• 제목/요약/키워드: Equine

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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
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    • 제57권5호
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    • pp.18.1-18.8
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    • 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.

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

  • 김민수;박종주;정윤희;이유연;;윤종택;민관식
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.221-221
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    • 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)

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

  • 이정희;정용제
    • 한국결정학회지
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    • 제5권1호
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    • pp.33-38
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    • 1994
  • Hen egg white Iysozyme과 equine serum albumin을 모델 단백질로 하여 'heterogeneous' vapor diffusion실험을 수행하였다. 즉, droplet과 reseroir용액에 각각 다른 침전제를 사용하여 vapor diffusion에 의하여 평형에 이르도록 하였다. 실험 결과는 NSI 혹은 ammonium sulfate 대신 polyethylene glycol이 reservoir 용액에 포함되므로써 droplet과 reservoir 용액 사이의 평형 속도가 감소되는 것을 보여 주었다. Heterogeneous vapor diffusion 법을 통하여 reservoir용액에 포함된 이온성 염과 비이온성 침전제의 양을 적절히 조절하여 평형속도를 조절할 수 있음을 보였다.

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