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莫祖英,车志群,廖思明,杨明柳,阎冰.多氯联苯胁迫下红树蚬荧光定量PCR内参基因的筛选[J].广西科学,2015,22(3):350-356. [点击复制]
- MO Zu-ying,CHE Zhi-qun,LIAO Si-ming,YANG Ming-liu,YAN Bing.Reference Gene Selection for qRT-PCR in Mangrove Mud Clam (Polymesoda erosa)Under PCBs Stress[J].Guangxi Sciences,2015,22(3):350-356. [点击复制]
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| 多氯联苯胁迫下红树蚬荧光定量PCR内参基因的筛选 |
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莫祖英1,2, 车志群2,3, 廖思明2,4, 杨明柳2, 阎冰2
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| (1.广西师范大学生命科学学院, 广西桂林 541004;2.广西红树林研究中心广西红树林保护与利用重点实验室, 广西北海 536007;3.广西大学生命科学技术学院, 广西南宁 530005;4.广西科学院广西生物科学与技术研究中心, 广西南宁 530007) |
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| 摘要: |
| [目的]筛选出多氯联苯(PCBs)胁迫下红树蚬(Polymesoda erosa)的实时荧光定量PCR (qRT-PCR)最适内参基因,为进一步开展分子毒理学研究奠定基础。[方法]以β-actin、18S rRNA、GAPDH和α-tubulin为候选内参基因,qRT-PCR测定PCBs胁迫下4个候选内参基因在红树蚬外套膜、鳃、闭壳肌和肝胰腺组织中的表达水平,应用geNorm、NormFinder和BestKeeper软件分析其表达稳定性。[结果]geNorm软件分析表明β-actin表达最稳定;NormFinder软件分析发现,外套膜、鳃和肝胰腺组织中β-actin的稳定性最好,闭壳肌组织中β-actin(0.454)表达稳定性略微低于α-tubulin(0.425),排第2位;BestKeeper软件分析表明,外套膜和鳃组织中β-actin最稳定,肝胰腺和闭壳肌组织中GAPDH最稳定,β-actin排位第2位。[结论]筛选β-actin为红树蚬在PCBs胁迫下的qRT-PCR最适内参基因。 |
| 关键词: 红树蚬 实时荧光定量PCR 内参基因 多氯联苯胁迫 |
| DOI： |
| 投稿时间：2015-03-04 |
| 基金项目:广西科学院基本科研业务费项目(13YJ22HS13)和广西红树林保护与利用重点实验室基金课题(GKLMC-08)资助。 |
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| Reference Gene Selection for qRT-PCR in Mangrove Mud Clam (Polymesoda erosa)Under PCBs Stress |
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MO Zu-ying1,2, CHE Zhi-qun2,3, LIAO Si-ming2,4, YANG Ming-liu2, YAN Bing2
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| (1.College of Life Sciences, Guangxi Normal University, Guilin, Guangxi, 541004, China;2.Guangxi Key Lab of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Beihai, Guangxi, 536007, China;3.College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;4.Guangxi Bioscience and Technology Research Center, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China) |
| Abstract: |
| [Objective] To select the suitable reference genes in the mangrove mud clam (Polymesoda erosa) exposed to polychlorinated biphenyls (PCBs) by using quantitative real-time PCR (qRT-PCR), which will be conducive to further research in molecular toxicology.[Methods] β-actin, 18S rRNA, GAPDH and α-tubulin were chosen as four candidate reference genes to test their expressed level in different tissues (mantle, gill, adductor muscle and hepatopancreas)of the mangrove mud clam under PCBs stress by using qRT-PCR method.The expression stability of these candidate reference genes were analyzed and evaluated with three software-based approaches (geNorm, NormFinder and BestKeeper).[Results] geNorm analysis showed that β-actin was the most stable expressed gene among the four candidate reference genes.NormFinder analysis found that β-actin was the most stable expressed gene in mantle, gill, and hepatopancreas, but in adductor muscle tissue, β-actin (0.454) was slightly less stable than α-tubulin (0.425) and ranked as the second.BestKeeper analysis showed that β-actin was the most stable expressed gene in mantle and gill tissues, whereas GAPDH was the most stable expressed gene and β-actin was ranked as the second in adductor muscle and hepatopancreas tissues.[Conclusion] β-actin can be used as a reference gene for mangrove mud clam under PCBs stress. |
| Key words: Polymesoda erosa quantitative real-time PCR reference genes polychlorinated biphenyls stress |
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