马氏珠母贝Su（H）基因克隆与组织特异性表达

摘要

Su（H）基因（Suppressor of Hairless gene）对生物体细胞的分化、增殖和凋亡起重要的调控作用。根据马氏珠母贝（Pinctada martensii）转录组数据库中注释为Su（H）的unigene序列设计基因特异性引物，应用cDNA末端快速扩增（RACE）技术克隆获得了马氏珠母贝Su（H）[pm-Su（H）]基因cDNA全长序列。结果表明：pm-Su（H）基因全长3830 bp，其中开放阅读框含有1920 bp，编码640个氨基酸残基，5'UTR为1568 bp，3'UTR为342 bp，含有27 bp polyA；预测其分子质量为70.6 ku，等电点为7.38；多序列比对显示，pm-Su （H）与其他物种的 Su（H）有较高的保守性，与牡蛎（Crassostrea gigas）的同源序列高达80%；荧光定量PCR分析表明，pm-Su（H）在马氏珠母贝闭壳肌、鳃、珍珠囊、外套膜、肝胰脏、性腺、足、血淋巴等8组织中均有表达，其中以性腺中表达量最高，其次是珍珠囊和外套膜。%Su (H) gene (Suppressor of Hairless) is conservative in evolution and plays an important role in cell differentiation, proliferation and apoptosis. Based on the unigene sequence annotated as Su(H) in the transcriptome database of Pinctada martensii, using rapid-amplification of cDNA ends (RACE) technology, the full length of Su(H) gene from Pinctada martensii [pm-Su(H)] was obtained. The full length of pm-Su(H) cDNA was 3 830 bp, with an open reading frame (ORF) of 1 920 bp, encoding 640 amino acid residues. The 3' untranslated region (UTR) was 1 568 bp and the 5' UTR was 342 bp. Multiple sequence alignment indicated that Su(H) was highly conservative among other species and pm-Su(H) are most closely associated with Crassostrea gigas. Meanwhile, quantitative real-time PCR analysis demonstrated that pm-Su(H) mRNA was constitutively expressed in adductor muscle, gill, pearl sac, mantle, hepatopancreas, gonad, foot and hemocyte, with the most abundant mRNA in the sexual gland.