高级检索+

巨龙竹秆形发育过程实时荧光定量PCR内参基因的筛选

Reference Gene Selection for Quantitative Real-Time PCR in Studying Culm Shape Development of Dendrocalamus sinicus

    摘要
  • 摘要:
    目的 利用荧光定量PCR(qRT-PCR)和表达分析软件筛选适用于巨龙竹秆形发育研究的qRT-PCR的稳定内参基因。
    方法 应用常规PCR和qRT-PCR, 分析EF-1α、GAPDH、Actin、Tubulin、TIP-41、PP2A共6个内参基因在不同秆形巨龙竹("通直型"和"弯曲型")竹笋3个不同发育时期的表达情况。利用GeNorm、NormFinder软件对各内参基因的表达稳定性进行评估; 以筛选的内参基因对巨龙竹木质素合成中的PTAL基因的表达量进行定量分析, 验证所筛选内参基因的有效性。
    结果 PCR和qRT-PCR的结果表明, 6个候选内参基因PCR目的片段电泳条带单一, 熔解曲线具有明显的单一峰。内参基因Actin、EF-1α、GAPDH的稳定性和相关性表现最佳; 以它们为内参, 对茎秆发育过程中PTAL基因进行定量分析, 结果显示它们均具有高效性。
    结论 Actin、EF-1α、GAPDH可以作为巨龙竹秆形发育研究中目的基因定量分析的内参基因。为进一步分析巨龙竹秆形发育过程中关键基因表达量的变化提供研究基础。

     

    Abstract:
    Objective To select suitable quantitative real-time PCR (qRT-PCR) reference genes for studying culm shape development of Dendrocalamus sinicus Chia et J. L. Sun.
    Method PCR and qRT-PCR were used to analyze the mRNA expression stability of six candidate reference genes (EF-1α, GAPDH, Actin, TIP-41, Tubulin and PP2A) in a set of six bamboo shoot samples at three development stages of the straight-culmed and bending-culmed D. sinicus respectively. The software GeNorm and NomFinder were employed to evaluate the data for reference genes. Quantitative analysis of PTAL was used to test the effectiveness of the candidate reference genes.
    Result The results showed that six candidate reference genes were of expected size and single peaking melting curve. Of them, Actin, EF-1α and GAPDH had the best stability and correlativity. Meanwhile, these three candidate reference genes had the highest efficiency based on quantitative analysis results of the PTAL gene during the culm development.
    Conclusion The Actin, EF-1α and GAPDH were optional inference genes for normalizing purpose genes in studying culm shape development of D. sinicus. The study can provide references for gene expression analysis in the culm development of D. sinicus.

     

    • HTML全文
    • 参考文献(24)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回