亚硝态氮胁迫下菲律宾蛤仔实时定量PCR内参基因的筛选
Alternative TitleSelection of Reference Genes by Quantitative Real-Time PCR for Ruditapes philippinarum Exposed to Nitrite
王爱云; 曹滕飞; 吕家森; 丛明
Source Publication生态毒理学报
ISSN1673-5897
2019
Volume14Issue:1Pages:153-160
Keyword亚硝态氮 菲律宾蛤仔 内参基因 转录组学 实时荧光定量pcr Nitrite Ruditapes Philippinarum Reference Gene Transcriptome Quantitative Real-time Pcr
MOST Discipline CatalogueEnvironmental Sciences & Ecology
Contribution Rank

(1) 鲁东大学生命科学学院

; (2) 烟台大学生命学院; (3) 中国科学院烟台海岸带研究所
Department海岸带环境过程实验室
Abstract

In order to well investigate the eco-toxicity of nitrite on molluscs, a suitable housekeeper gene was needed for further analysis to qualify the toxicity-related genes. In the present study, the clam Ruditapes philippinarum was used as the target mollusc and cDNAs templates were got from gill tissue RNA of R. philippinarum after nitrite exposure. Six candidate genes were assayed in terms of their expression stabilities by geNorm and NormFinder softwares, followed by a comprehensive weighting comparison. GeNorm analysis indicated that the stability order for the six genes were Actin>CyPA>EF1alpha>Ubi>18S>Tubu. However, a little different order was suggested by the Normfinder software, Actin>Ubi>CyPA>EF1alpha>18S>Tubu. After a comprehensive weighting comparison, the final stability order of the six genes was Actin>CyPA>Ubi>EF1alpha>18S>Tubu. So, the Actin gene was the most suitable reference gene to qualify target genes for their expression levels in the clam R. philippinarum after nitrite exposure.

Other Abstract

为了更好地从基因角度研究亚硝态氮对贝类的生态毒性,需要筛选一个合适的内参基因作为参考,对其毒性相关基因进行定量表达分析。本研究以菲律宾蛤仔(Ruditapes philippinarum)作为研究对象,以亚硝态氮胁迫的鳃组织cDNA作为模板,利用geNorm软件和NormFinder软件对6个候选内参基因进行表达稳定性分析,并对2个结果做了综合赋权分析。结果显示, geNorm分析的基因表达稳定性结果为Actin>CyPA>EF1alpha>Ubi>18S>Tubu,NormFinder软件分析的结果为Actin>Ubi>CyPA> EF1alpha>18S>Tubu,最后经加权赋值法综合分析获得6个候选基因的表达稳定性为Actin>CyPA>Ubi>EF1alpha>18S>Tubu,即Actin为表达最稳定的基因。因此,Actin可以作为衡量亚硝态氮胁迫后菲律宾蛤仔鳃组织中基因表达的内参基因。

Indexed ByCSCD
Language中文
WOS Keyword亚硝态氮 ; 菲律宾蛤仔 ; 内参基因 ; 转录组学 ; 实时荧光定量pcr ; Nitrite ; Ruditapes Philippinarum ; Reference Gene ; Transcriptome ; Quantitative Real-time Pcr
WOS Research AreaEnvironmental Sciences & Ecology
CSCD IDCSCD:6506667
Citation statistics
Document Type期刊论文
Identifierhttp://ir.yic.ac.cn/handle/133337/25154
Collection中科院海岸带环境过程与生态修复重点实验室_海岸带环境过程实验室
Affiliation1.鲁东大学生命科学学院;
2.烟台大学生命学院;
3.中国科学院烟台海岸带研究所
Recommended Citation
GB/T 7714
王爱云,曹滕飞,吕家森,等. 亚硝态氮胁迫下菲律宾蛤仔实时定量PCR内参基因的筛选[J]. 生态毒理学报,2019,14(1):153-160.
APA 王爱云,曹滕飞,吕家森,&丛明.(2019).亚硝态氮胁迫下菲律宾蛤仔实时定量PCR内参基因的筛选.生态毒理学报,14(1),153-160.
MLA 王爱云,et al."亚硝态氮胁迫下菲律宾蛤仔实时定量PCR内参基因的筛选".生态毒理学报 14.1(2019):153-160.
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