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Vol. 64 No. 12: Issue 12

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Investigation on the expression stability of common reference genes in Aurelia sp.1 under hypoxia

https://doi.org/10.14715/cmb/2018.64.12.6
Xiang Gao
College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, P. R. China2. Key Laboratory of Marine Environment and Ecology, Ministry of Education, 238 Songling Road, Qingdao 266100, P. R. China
Guoshan Wang
College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, P. R. China2. Key Laboratory of Marine Environment and Ecology, Ministry of Education, 238 Songling Road, Qingdao 266100, P. R. China

Corresponding Author(s) : Guoshan Wang

vs1290@163.com

Cellular and Molecular Biology, Vol. 64 No. 12: Issue 12

Abstract

RT-qPCR (Quantitative real-time polymerase chain reaction) is a reliable molecular biology technique used for gene expression detection due to its high sensibility and good reproducibility. However, suitable reference genes for RT-qPCR are often not available to investigate the expression of target genes in jellyfish under different conditions. To determine the responsible genes of jellyfish under hypoxia, primers to amplify the actin gene was designed for the amplification according to the conserved actin amino acid sequences of cnidarian. Then, we cloned and sequenced the partial cDNA sequence of β-actin gene containing 849 bp nucleic acids was cloned and sequenced, and the four common housekeeping genes (18S rRNA, β-actin, α-tubulin and GAPDH) were detected. To obtain suitable reference genes, we compared the four genes under normoxia and hypoxia were determined and compared using RT-qPCR. The evaluation result shows that α-tubulin gene can be used as single reference gene, and α-tubulin and β-actin can be served as multiple reference genes to study relative gene expression related to hypoxic tolerance of Aurelia sp.1. This research will establish foundation to reveal the molecular mechanism of jellyfish under hypoxia.

Keywords

Aurelia sp.1 β-actin Reference gene RFQ-PCR.
Gao, X., & Wang, G. (2018). Investigation on the expression stability of common reference genes in Aurelia sp.1 under hypoxia. Cellular and Molecular Biology, 64(12), 26–31. https://doi.org/10.14715/cmb/2018.64.12.6
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References
  1. Sun S, Sun XX, Jenkinson IR. Preface: Giant jellyfish blooms in Chinese waters. Hydrobiologia 2015; 754: 1-11.
  2. Wu L, Wang J, Gao S, Zheng X, Huang R. An analysis of dynamical factors influencing 2013 giant jellyfish bloom near Qinhuangdao in the Bohai Sea, China ∗. Estuar Coast Shelf Sci 2016; 185: 10.
  3. Lingjuan WU, Xiaofen WU, Bai T. Comprehensive analysis of the origin of giant jellyfish near Qinhuangdao in summer. Chin J Oceanol Limnol 2017; 35: 1-9.
  4. Zhang F, Sun S, Jin X, Li C. Associations of large jellyfish distributions with temperature and salinity in the Yellow Sea and East China Sea. Hydrobiologia 2012; 690: 81-96.
  5. Dong J, Sun M, Purcell JE, Chai Y, Zhao Y, Wang A. Effect of salinity and light intensity on somatic growth and podocyst production in polyps of the giant jellyfish Nemopilema nomurai (Scyphozoa: Rhizostomeae). Hydrobiologia 2015; 754: 75-83.
  6. Dong Z, Liu D, Keesing JK. Jellyfish blooms in China: Dominant species, causes and consequences. Mar Pollut Bull 2010; 60: 954-963.
  7. Kim DH, Seo JN, Yoon WD, Suh YS. Estimating the economic damage caused by jellyfish to fisheries in Korea. Fish Sci 2012; 78: 1147-1152.
  8. Shoji J, Kudoh T, Takatsuji H, Kawaguchi O, Kasai A, Chen ZY, et al. Distribution of moon jellyfish Aurelia aurita in relation to summer hypoxia in Hiroshima Bay, Seto Inland Sea. Estuar Coast Shelf Sci 2010; 86: 485-490.
  9. Miller MEC, Graham WM. Environmental evidence that seasonal hypoxia enhances survival and success of jellyfish polyps in the northern Gulf of Mexico. J Exp Mar Biol Ecol 2012; 432: 113-120.
  10. Wang G, Yu Z, Zhen Y, Mi T, Shi Y, Wang J, et al. Molecular Characterisation, Evolution and Expression of Hypoxia-Inducible Factor in Aurelia sp.1. Plos One 2014; 9: 100057.
  11. Huggett J, Dheda K, Bustin S, Zumla A. Real-time RT-PCR normalisation; strategies and considerations. Genes Immun 2005; 6: 279-284.
  12. Yang G, Bao B, Ren D. Comparison between ribosomal 18S rRNA,GAPDH and #beta#-actin genes as internal standard for quantitative comparison of mRNA levels in development of flounder,Paralichthys olivaceus. J Shanghai Fish Univ 2005; 14: 84-88.
  13. Kim BR, Nam HY, Kim SU, Kim SI, Chang YJ. Normalization of reverse transcription quantitative-PCR with housekeeping genes in rice. Biotechnol Lett 2003; 25: 1869-1872.
  14. Gutierrez N, Giménez MJ, Palomino C, Avila CM. Assessment of candidate reference genes for expression studies in Vicia faba L. by real-time quantitative PCR. Mol Breed 2011; 28: 13-24.
  15. Park SJ, Kim YH, Lee Y, Kim KM, Kim HS, Lee SR, et al. Selection of Appropriate Reference Genes for RT-qPCR Analysis in a Streptozotocin-Induced Alzheimer's Disease Model of Cynomolgus Monkeys (Macaca fascicularis). Plos One 2013; 8: 56034.
  16. Gu C, Chen S, Liu Z, Shan H, Luo H, Guan Z, et al. Reference gene selection for quantitative real-time PCR in Chrysanthemum subjected to biotic and abiotic stress. Mol Biotechnol 2011; 49: 192-197.
  17. Lin YLL, Z X. Reference gene selection for qPCR analysis during somatic embryogenesis in longan tree. Plant Sci 2010; 178: 359-365.
  18. Selvey S, Thompson EW, Matthaei K, Lea RA, Irving MG, Griffiths LR. Beta-actin--an unsuitable internal control for RT-PCR. Mol Cell Probes 2001; 15: 307-311.
  19. Rytkönen KT, Renshaw GM, Ashton KJ, Williamspritchard G, Leder EH, Nikinmaa M. Elasmobranch qPCR reference genes: a case study of hypoxia preconditioned epaulette sharks. BMC Mol Biol 2010; 11: 27.
  20. Andersen CL, Jensen JL, í˜rntoft TF. Normalization of Real-Time Quantitative Reverse Transcription-PCR Data: A Model-Based Variance Estimation Approach to Identify Genes Suited for Normalization, Applied to Bladder and Colon Cancer Data Sets. Cancer Res 2004; 64: 5245.
  21. Vandesompele J, Preter KD, Pattyn F, Poppe B, Roy NV, Paepe AD, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002; 3: 34.
  22. Zhu J, Zhang L, Li W, Han S, Yang W, Qi L. Reference Gene Selection for Quantitative Real-time PCR Normalization in Caragana intermedia under Different Abiotic Stress Conditions. Plos One 2013; 8: 53196.
  23. Teng X, Zhang Z, He G, Yang L, Li F. Validation of reference genes for quantitative expression analysis by real-time RT-PCR in four lepidopteran insects. J Insect Sci 2012; 12: 1-17.
  24. Wittkopp PJ, Kalay G. Cis-regulatory elements: molecular mechanisms and evolutionary processes underlying divergence. Nat Rev Genet 2012; 13: 59-69.
  25. Ereskovsky AV, Renard E, Borchiellini C. Cellular and molecular processes leading to embryo formation in sponges: evidences for high conservation of processes throughout animal evolution. DDev Genes Evol 2013; 223: 5.
  26. Bridge D, Cunningham CW, Schierwater B, Desalle R, Buss LW. Class-level relationships in the phylum Cnidaria: evidence from mitochondrial genome structure. Proc Natl Acad Sci U S A 1992; 89: 8750-8753.
  27. Collins AG. Phylogeny of Medusozoa and the evolution of cnidarian life cycles. J Evol Biol 2002; 15: 418-432.
  28. Chi X, Hu R, Yang Q, Zhang X, Pan L, Chen N, et al. Validation of reference genes for gene expression studies in peanut by quantitative real-time RT-PCR. Mol Genet Genom 2012; 287: 167-176.
  29. Suzuki T, Higgins PJ, Crawford DR. Control selection for RNA quantitation. Biotechniques 2000; 29: 332-337.
  30. Higashimura Y, Nakajima Y, Yamaji R, Harada N, Shibasaki F, Nakano Y, et al. Up-regulation of glyceraldehyde-3-phosphate dehydrogenase gene expression by HIF-1 activity depending on Sp1 in hypoxic breast cancer cells. Arch Biochem Biophys 2011; 509: 1.
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