大菱鲆脾脏cDNA文库构建、EST序列分析与免疫抗病相关基因的筛选

Construction of cDNA library from spleen of Scophthalmus maximus and identification of immune-related genes by EST sequencing

以大菱鲆脾脏为材料,构建cDNA文库,通过对文库克隆的序列测定和初步生物信息学分析,共获得3 656个大菱鲆脾脏表达序列标签(EST)。经与GenBank数据库的序列比对后发现,1 891个EST代表了149个已知基因,其余1 765个EST为未知序列。这149个基因大致可分为6类:9个为细胞结构类(6.0%);26个为代谢类(17.5%);45个为细胞防御/免疫类(30.2%);43个为基因/蛋白质表达类(28.9%);16个为细胞信号转导/细胞交流类(10.7%);10个无法明确分类(6.7%)。鉴定的与免疫抗病相关重要基因主要有:CC/CXC趋化因子;主要组织相容性复合体(MHC)class I;αC3补体;天然杀伤细胞增强因子(NKEF);胸腺素β-4/β-12;干扰素诱导蛋白G ig1;白介素8受体;STAT4;热激蛋白70/90等。

A cDNA library was constructed from turbot, Scophthalmus maximus, spleen by unidirectional cloning. A total of 3 656 ESTs from the library were sequenced and compared with sequences in the GenBank database. 1 765 clones （48.3%） appeared to be completely unknown and are likely to represent newly described genes, whereas 1 891 clones （51.7%） were identified based on matches to sequences in the database. These identified clones were derived from at least 149 genes which were categorized into six categories： 9 in cell structure/motility （6.0%）, 26 in metabolism （17.5%）, 45 in cell defense/immunity （30. 2% ）, 43 in gene/protein expression （28. 9% ）, 16 in cell signal transduction/communication （10.7%）, and 10 genes lacking enough information to be classified （6.7%）. Immune related cDNAs identified from the spleen were： CC/CXC chemokine, MHC class Iα, complement component C1/C3, natural killer cell enhancing factor （NKEF）, thymosin beta-4/beta-12, interferon inducible protein Gigl, IL-B4 receptor, STAT4, and hot shock protein 70/90.