Identification of normalization factors for quantitative realtime RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai
文献类型：CNKI期刊论文
| 作者 | Qiu R(邱礽); Sun BG(孙铂光) ; Fang SS(房沙沙); Sun L(孙黎); Liu X(刘晓)
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| 发表日期 | 2013-03-15 |
| 出处 | Chinese Journal of Oceanology and Limnology
 |
| 关键词 | Haliotis discus hannai housekeeping gene normalization factor quantitative real time RT-PCR reference gene |
| 英文摘要 | Quantitative real-time reverse transcription-polymerase chain reaction(qRT-PCR) is widely used in studies of gene expression.In most of these studies,housekeeping genes are used as internal references without validation.To identify appropriate reference genes for qRT-PCR in Pacific abalone Haliotis discus hannai,we examined the transcription stability of six housekeeping genes in abalone tissues in the presence and absence of bacterial infection.For this purpose,abalone were infected with the bacterial pathogen Vibrio anguillarum for 12 h and 48 h.The mRNA levels of the housekeeping genes in five tissues(digestive glands,foot muscle,gill,hemocyte,and mantle) were determined by qRT-PCR.The PCR data was subsequently analyzed with the geNorm and NormFinder algorithms.The results show that in the absence of bacterial infection,elongation factor-1-alpha and beta-actin were the most stably expressed genes in all tissues,and thus are suitable as cross-tissue type normalization factors.However,we did not identify any universal reference genes post infection because the most stable genes varied between tissue types.Furthermore,for most tissues,the optimal reference genes identified by both algorithms at 12 h and 48 h post-infection differed.These results indicate that bacterial infection induced significant changes in the expression of abalone housekeeping genes in a manner that is dependent on tissue type and duration of infection.As a result,different normalization factors must be used for different tissues at different infection points. |
| 文献子类 | CNKI期刊论文 |
| 资助机构 | Supported by the Knowledge Innovation Program of Chinese Academy of Sciences(No.KSCX2-EW-G-12B) ; the Knowledge Innovation Program of the Chinese Academy of Sciences(No.KZCX2-EW-Q213) ; the National High Technology Research and Development Program of China (863 Program)(No.2012AA10A412) |
| 卷 | v.31期:02页:177-186 |
| 语种 | 英文; |
| 分类号 | S917.4 |
| ISSN号 | 0254-4059 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/190933]  |
| 专题 | 中国科学院海洋研究所 |
| 作者单位 | 1.GraduateUniversityofChineseAcademyofSciences 2.InstituteofOceanology,ChineseAcademyofSciences |
| 推荐引用方式 GB/T 7714 | Qiu R,Sun BG,Fang SS,et al. Identification of normalization factors for quantitative realtime RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai. 2013. |
入库方式： OAI收割
来源：海洋研究所
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