SELECTION OF INTERNAL CONTROL GENE FOR EXPRESSION STUDIES IN PORPHYRA HAITANENSIS (RHODOPHYTA) AT DIFFERENT LIFE-HISTORY STAGES(1)
Xiaojie Wu1, Jianfeng Niu1, Aiyou Huang1
1College of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaKey Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences (IOCAS), Qingdao 266071, China Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences (IOCAS), Qingdao 266071, China Graduate University of the Chinese Academy of Sciences, Beijing 100049, ChinaCollege of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China Key Laboratory of Experimental Marine Biology, Institute of Oceanology, the Chinese Academy of Sciences (IOCAS), Qingdao 266071, China College of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China.
Abstract:
Accurate gene quantification depends on the use of an appropriate internal control gene, which should be verified before its use for normalizing data. Housekeeping genes, which are expressed at relatively constant levels, are generally regarded as candidate internal control genes. To determine the ideal internal control for gene expression profiles for Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) at different life-history stages, we used absolute quantification to assess the expression levels of six housekeeping genes (18S ribosomal RNA, 30S ribosomal protein, glyceraldehyde-3-phosphate dehydrogenase, elongation factor 3, alpha-tubulin, and beta-tubulin) at the sporophyte and gametophyte stages. Housekeeping genes were selected by comparing the differences of observed copy numbers in sporophytes and in gametophytes. TubB (beta-tubulin) was found to be the optimal internal control gene, because it showed the smallest difference of gene expression. Compared with TubB, other housekeeping genes had greater variation of expression to different degrees.
