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      Validation of Housekeeping Genes for Gene Expression Analysis in Iwagaki Oyster ( Crassostrea nippona) Under Salinity Stress by Quantitative Real-Time PCR

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          Abstract

          Hypo-salinity can reduce the immunological reaction in Crassostrea nippona, even lead to massive mortality. It is important to understand the molecular mechanism of oyster defense system, while quantitative real-time PCR can be employed in the study. However, the accuracy of quantitative real-time PCR relies on the use of suitable reference genes. In this study, the expression stability of 14 candidate reference genes including traditional housekeeping genes EF1A, TUB, TUA, GAPDH, RO21, as well as new candidate reference genes RPL5, RPL8, RPS27, RPL14, RPL4, CO3, RPS8, RPS4, CYTB in different tissues of C. nippona under salinity stress has been validated by quantitative real-time PCR. Ribosomal protein genes selected through expression analysis of transcriptome data from C. nippona generally were more stable than traditional reference genes. According to the geNorm analysis, RPL4 and RPS4 could be used as internal controls for studying gene expression in C. nippona with real-time PCR under salinity stress.

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          Author and article information

          Journal
          JOUC
          Journal of Ocean University of China
          Science Press and Springer (China )
          1672-5182
          15 November 2020
          01 December 2020
          : 19
          : 6
          : 1441-1446
          Affiliations
          1Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China
          2Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China
          Author notes
          *Corresponding author: LI Qi, Tel: 0086-532-82031622, E-mail: qili66@ 123456ouc.edu.cn
          Article
          s11802-020-4422-4
          10.1007/s11802-020-4422-4
          Copyright © Ocean University of China, Science Press and Springer-Verlag GmbH Germany 2020.

          The copyright to this article, including any graphic elements therein (e.g. illustrations, charts, moving images), is hereby assigned for good and valuable consideration to the editorial office of Journal of Ocean University of China, Science Press and Springer effective if and when the article is accepted for publication and to the extent assignable if assignability is restricted for by applicable law or regulations (e.g. for U.S. government or crown employees).

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          Self URI (journal-page): https://www.springer.com/journal/11802

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