家蚕Osiris基因家族的克隆与序列结构及表达研究  

Cloning,Sequence Structure and Expression Analysis of Osiris Gene Family in the Silkworm,Bombyx mori

作　　者：王更先[1,2] 司马杨虎[1] 张升祥[3] 徐世清[1] 

机构地区：[1]苏州大学现代丝绸国家工程实验室苏州大学基础医学与生物科学学院,江苏苏州215123 [2]邯郸学院生物科学系,河北邯郸056005 [3]山东农业大学林学院,山东泰安271018

出　　处：《华北农学报》2010年第6期1-8,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB121005)

摘　　要：利用生物信息学、RT-PCR和RACE等方法克隆了家蚕6个Osiris基因家族的成员,依次命名为BmOsiris7、BmOsiris9、BmOsiris18、BmOsiris19、BmOsiris20和BmOsiris21。序列分析表明,BmOsiris基因的序列ORF长度在723~882之间,所编码的氨基酸分子量大小也很接近,在26~31 kDa之间。pI分析表明,BmOsiris21为9.10,其余在6.41~8.66之间。SMART在线保守区域预测发现,该蛋白家族属于跨膜蛋白,都含有Osiris家族特有的DUF1676保守区域,支持了克隆的所有基因序列属于Osiris基因家族的推测。Osiris家族蛋白序列进化树分析显示,家蚕和黑腹果蝇的同系同源蛋白要近于同物种的旁系同源蛋白。表达谱分析表明,各基因在所调查的组织中均没有组织和时空特异性,但表达量的多少存在显著差异。染色体定位结果显示,BmOsiris7、BmOsiris9、BmOsiris18、BmOsiris19和BmOsiris20定位在Chr.26上,而BmOsiris 21单独定位在了Chr.4上。家蚕Osiris基因家族的以上所有分析结果和黑腹果蝇中O-siris基因家族极其相似,推测家蚕Osiris家族的功能可能与果蝇相似。Triplo-lethal Locus（Tpl） was surveyed as the only one that both triplo-lethal and haplo-lethal gene locus in the Drosophila genome.There was an Osiris gene family named gene Osiris1 through Osiris20 in the Tpl region.Through BLASTTP and PSI-BLAST analysis,there were only three other members of this family located outside of this locus.We called them Osiris21 to Osiris23.This gene family had the Insecta specificity.Through bioinformatics,RT-PCR and RACE methods,six silkworm genes belonging to Osiris gene family have been cloned and named as BmOsiris7,BmOsiris9,BmOsiris18,BmOsiris19,BmOsiris20 and BmOsiris21,respectively.Sequence analysis indicated that BmOsiris gene′s ORF were between 728 bp and 882 bp in length.The molecular size of deduced amino acid which seemed quite similar with Drosophila Osiris protein and were 26-31 kDa range.The analysis of isoelectric point indicated that BmOsiris21′s isoelectric point was 9.0,others were at between 6.41 and 8.66.SMART conservative region prediction on line found that this protein family belong to transmembrane protein and had specific conservative regions for the Osiris family,called DUF1676.All above dated supported the speculation that the cloned genes belonged to Osiris gene family.Phylogenetic tree analysis demonstrated paralogs of Bombyx mori and Drosophila melanogaster were much closer to the infraspecific orthologs.Expression pattern analysis indicated that each gene in our tested samples had no tissue and space-time speciality,but expression amount had significant differences.chromosome location showed that BmOsiris7、BmOsiris9、 BmOsiris18、BmOsiris19 and BmOsiris20 located on the Chr.26,however,BmOsiris21 individually located on Chr.4.This result was very similar with Osiris genes from Drosophila melanogaster.

关 键 词：家蚕 Osiris基因家族 基因克隆 序列分析 基因表达 

分 类 号：S881.2[农业科学—特种经济动物饲养] S186[农业科学—畜牧兽医]