Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis
文献类型：CNKI期刊论文
| 作者 | Wang X(王霞) ; Feng JH(冯建华); Huang AY(黄爱优); He LW(何林文); Niu JF(牛建峰); Wang GC(王广策)
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| 发表日期 | 2017-11-15 |
| 出处 | Chinese Journal of Oceanology and Limnology
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| 关键词 | real-time quantitative PCR housekeeping genes internal control genes stress responding Pyropia haitanensis |
| 英文摘要 | Pyropia haitanensis has prominent stress-resistance characteristics and is endemic to China. Studies into the stress responses in these algae could provide valuable information on the stress-response mechanisms in the intertidal R hodophyta. Here, the ef fects of salinity and light intensity on the quantum yield of photosystem II in P y. haitanensis were investigated using pulse-amplitude-modulation fluorometry. Total RNA and genomic DNA of the samples under dif ferent stress conditions were isolated. By normalizing to the genomic DNA quantity, the RNA content in each sample was evaluated. The c DNA was synthesized and the expression levels of seven potential internal control genes were evaluated using q RT-PCR method. Then, we used ge Norm, a common statistical algorithm, to analyze the q RT-PCR data of seven reference genes. Potential genes that may constantly be expressed under dif ferent conditions were selected, and these genes showed stable expression levels in samples under a salinity treatment, while t ubulin, glyceraldehyde-3-phosphate dehydrogenase and actin showed stability in samples stressed by strong light. Based on the results of the pulse amplitude-modulation fluorometry, an absolute quantification was performed to obtain gene copy numbers in certain stress-treated samples. The stably expressed genes as determined by the absolute quantification in certain samples conformed to the results of the ge Norm screening. Based on the results of the software analysis and absolute quantification, we proposed that e longation factor 3 and 18 S ribosomal RNA could be used as internal control genes when the P y. haitanensis blades were subjected to salinity stress, and that α-tubulin and 18 S ribosomal RNA could be used as the internal control genes when the stress was from strong light. In general, our findings provide a convenient reference for the selection of internal control genes when designing experiments related to stress responses in Py. haitanensis. |
| 文献子类 | CNKI期刊论文 |
| 资助机构 | Supported by the National Natural Science Foundation of China(Nos.41476140,41306151,41676157,41506172) ; the Strategic Leading Science and Technology Projects of Chinese Academy of Sciences(No.XDA11020404) ; the China Postdoctoral Science Foundation(No.2015M582153) ; the Science and Technology Plan of Jiangsu Province(No.BE2016330) |
| 卷 | v.35期:06页:174-183 |
| 语种 | 英文; |
| 分类号 | Q943.2 |
| ISSN号 | 0254-4059 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/189089]  |
| 专题 | 中国科学院海洋研究所 |
| 作者单位 | 1.LaboratoryforMarineBiologyandBiotechnology,QingdaoNationalLaboratoryforMarineScienceandTechnology 2.KeyLaboratoryofExperimentalMarineBiology,InstituteofOceanology,ChineseAcademyofSciences 3.NantongBranch,InstituteofOceanology,ChineseAcademyofSciences 4.UniversityofChineseAcademyofSciences |
| 推荐引用方式 GB/T 7714 | Wang X,Feng JH,Huang AY,et al. Identification of potential internal control genes for real-time PCR analysis during stress response in Pyropia haitanensis. 2017. |
入库方式： OAI收割
来源：海洋研究所
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