基于抑制消减杂交识别白化刺参和青刺参差异表达基因

Identification of Differentially Expressed Genes Between Albino and Normal Apostichopus japonicus

为探寻刺参体色遗传的内在分子基础,运用Clontech SSH试剂盒分别构建了白化刺参和青刺参的正反向抑制性消减杂交文库;221个克隆测序分析确认了体色遗传差异性表达基因26个;GO分析显示这些差异基因分别隶属于核糖体基因、能量代谢、细胞结构、免疫调控、白化性状直接相关基因及未知基因.荧光定量PCR结果显示:ADP-核糖激化因子在白化刺参中相对表达量约是青刺参的2倍,而虾青素和小眼畸形转录因子在白化刺参中相对表达量则低于青刺参,分别为青刺参的0.38倍和0.47倍,相对表达量趋势和文库所得结果一致.该研究为进一步了解刺参体色分化及其优良性状的形成机制提供了基础资料.

To explore the intrinsic molecular issues behind the different growth and resistance, we construct a forward and reverse suppression subtractive hybridization database using albino and normal sea cucumbers. GO analysis indicates that 26 differentially expressed genes are classified into six categories, including ribosomal genes, metabolism related genes cell structure, immune gene, albino related genes and unknown genes. RT-qPCR analysis shows that the relative expression of ADP-ribosylation factor in albino sea cucumber is twice as much as that of normal ones. On the contrary, the relative expression of astacin and microphthalmia-associated transcription factor in albino sea cucumber is 2.6 and 2.1 times higher than normal ones, which is consistent with those recorded in SSH library. The present study provides the basic insight for further understanding the mechanism of eolour differentiation for sea cucumbers.