Validation of housekeeping genes as internal controls for studying the gene expression in Pyropia ha

来源 :Acta Oceanologica Sinica | 被引量 : 0次 | 上传用户:ljc20090204
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Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. To explore the mechanisms underlying these characteristics, gene expression has been investigated at the whole transcriptome level. Gene expression studies using quantitative real-time PCR should start by selecting an appropriate internal control gene; therefore, the absolute expression abundance of six housekeeping genes(18S rRNA(18S), ubiquitin-conjugating enzyme(UBC), actin(ACT), β-tubulin(TUB), elongation factors 2(EF2), and glyceraldehyde-3-phosphate dehydrogenase(GAPDH) examined by the quantitative real-time PCR in samples corresponding to different strains, life-cycle stages and abiotic stress treatments. Their expression stabilities were assessed by the comparative cycle threshold(Ct) method and by two different software packages: geNorm and NormFinder. The most stable housekeeping gene is UBC and the least stable housekeeping is GADPH. Thus, it is proposed that the most appropriate internal control gene for expression analyses in P. haitanensis is UBC. The results pave the way for further gene expression analyses of different aspects of P. haitanensis biology including different strains, life-history stages and abiotic stress responses. Pyropia haitanensis is an economically important mariculture crop in China and has a high research value for several life phenomena, for example environmental tolerance. For example environmental expression. Gene expression studies has been investigated at the whole transcriptome level. The real-time PCR should start with selecting an appropriate internal control gene; therefore, the absolute expression abundance of six housekeeping genes (18S rRNA (18S), ubiquitin-conjugating enzyme (UBC), actin , elongation factors 2 (EF2), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) examined by the quantitative real-time PCR in samples corresponding to different strains, life-cycle stages and abiotic stress treatments. Their expression stabilities were assessed by the comparative cycle threshold (Ct) method and by two different software packages: geNorm and NormFinder. The most stable housekeeping gene is UBC and the least st Thus, it is proposed that the most appropriate internal control gene for expression analyzes in P. haitanensis is UBC. The results pave the way for further gene expression analyzes of different aspects of P. haitanensis biology including different strains, life -history stages and abiotic stress responses.
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