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Validation of reference genes for gene expression studies in the dinoflagellate Akashiwo sanguinea by quantitative real-time RT-PCR
  • ISSN号:0253-505X
  • 期刊名称:《海洋学报:英文版》
  • 分类:P[天文地球]
  • 作者机构:[1]Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China, [2]Laboratory for Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China, [3]University of the Chinese Academy of Sciences, Beijing 100049, China
  • 相关基金:The National Natural Science Foundation of China-Shandong Joint Fund for Marine Science Research Centers under contract No.U1406403; the State Key Program of National Natural Science of China under contract No.61533011; China Postdoctoral Science Foundation under contract Nos 2014M551969 and 2015T80754
中文摘要:

The accurate measurement of gene expression via quantitative real-time reverse transcription PCR(q RT-PCR)heavily relies on the choice of valid reference gene(s) for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA(18S), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), α-tubulin(TUA), β-tubulin(TUB), actin(ACT) and cytochrome oxidase subunit 1(COX1), were evaluated for their expression stability with q RT-PCR and three statistical algorithms(Ge Norm, Norm Finder, and Best Keeper) for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages(resting cysts), different growth stages, in darkness, exposed to abscisic acid(ABA) and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates regarding quantification of gene expression a

英文摘要:

The accurate measurement of gene expression via quantitative real-time reverse transcription PCR(q RT-PCR)heavily relies on the choice of valid reference gene(s) for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA(18S), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), α-tubulin(TUA), β-tubulin(TUB), actin(ACT) and cytochrome oxidase subunit 1(COX1), were evaluated for their expression stability with q RT-PCR and three statistical algorithms(Ge Norm, Norm Finder, and Best Keeper) for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages(resting cysts), different growth stages, in darkness, exposed to abscisic acid(ABA) and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates r

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期刊信息
  • 《海洋学报:英文版》
  • 主管单位:
  • 主办单位:中国海洋学会
  • 主编:潘德炉
  • 地址:北京海淀大慧寺路8号
  • 邮编:100081
  • 邮箱:hyxbe@263.net
  • 电话:010-62179976
  • 国际标准刊号:ISSN:0253-505X
  • 国内统一刊号:ISSN:11-2056/P
  • 邮发代号:82-285
  • 获奖情况:
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),荷兰文摘与引文数据库,美国剑桥科学文摘,美国科学引文索引(扩展库),英国科学文摘数据库,英国动物学记录
  • 被引量:331