| 引用本文： |
-
车志群,孙仁杰,刘文爱,廖思明,阎冰.六溴环十二烷胁迫下红树蚬荧光定量PCR内参基因稳定性分析[J].广西科学院学报,2018,34(3):242-249. [点击复制]
- CHE Zhiqun,SUN Renjie,LIU Wen'ai,LIAO Siming,YAN Bing.The Expression Stability Analysis of Reference Genes for qRT-PCR in Polymesoda erosa under Hexabromocyclododecane Stress[J].Journal of Guangxi Academy of Sciences,2018,34(3):242-249. [点击复制]
|
|
| 摘要: |
| [目的]筛选出不同浓度六溴环十二烷(HBCD)胁迫下红树蚬(Polymesoda erosa)实时荧光定量PCR (qRT-PCR)的最适内参基因,用以准确校准目的基因的表达,为进一步开展分子毒理研究和开发分子生物标志物奠定基础。[方法]通过荧光定量PCR技术监测候选内参18S核糖体RNA (18S rRNA)、β-肌动蛋白(β-actin)、甘油醛-3-磷酸脱氢酶(GAPDH)、α-微管蛋白(α-tubulin)等管家基因,在红树蚬各组织受不同浓度(0 μg/L、0.86 μg/L、8.6 μg/L)六溴环十二烷(HBCD)胁迫后的表达量,利用qRT-PCR及geNorm、NormFinder、BestKeeper等分析软件对不同条件下的表达数据进行处理,从而筛选出适合荧光定量PCR的最佳内参基因。[结果]受不同浓度HBCD胁迫后,鳃及肝胰中各管家基因的表达稳定性排序整体为β-actin=α-tubulin > GAPDH > 18S rRNA。[结论]可单独或共同使用两个内参基因(β-actin、α-tubulin)校准荧光定量结果。 |
| 关键词: 红树蚬 六溴环十二烷 内参基因 稳定性分析 |
| DOI：10.13657/j.cnki.gxkxyxb.20180807.001 |
| 投稿时间：2018-03-20 |
| 基金项目:广西红树林保护与利用重点实验室开放基金(GKLMC-201306),广西红树林保护与利用重点实验室基金课题(GMLMC-201310)和科技部林业公益项目(201504413)资助。 |
|
| The Expression Stability Analysis of Reference Genes for qRT-PCR in Polymesoda erosa under Hexabromocyclododecane Stress |
|
CHE Zhiqun1,2, SUN Renjie1, LIU Wen'ai1, LIAO Siming3, YAN Bing1
|
| (1.Guangxi Key Lab of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Guangxi Academy of Sciences, Beihai, Guangxi, 536000, China;2.Guangxi University, Nanning, Guangxi, 530000, China;3.Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China) |
| Abstract: |
| [Objective] This experiment was to select the best reference gene for quantitative PCR under different concentrations of hexabromocyclododecane treatments, which will accurately calibrate the expression of objective genes and lay a foundation for further study on molecular toxicology research and development of molecular biomarkers. [Methods] The expression level of four candidate genes(18S rRNA、β-actin、GAPDH、α-tubulin)on different tissues of Polymesoda erosa under different concentrations of hexabromocyclododecane treatments were monitored by using qRT-PCR technology, and the expression data obtained from qRT-PCR was analyzed with different software, such as BestKeeper, geNorm, NormFinder to select the best reference gene suitable for real-time PCR. [Results] The results showed that after being stressed by different concentrations of HBCD, the order of candidate genes expression in gill and hepatopancreas was:β-actin=α-tubulin > GAPDH > 18S rRNA. [Conclusion] The qRT-PCR results can be calibrated using two reference genes (β-actin, α-tubulin) alone or in combination. |
| Key words: Polymesoda erosa hexabromocyclododecane reference genes stability analysis |
