利用mRNA差异显示技术筛选丝羽乌骨鸡肝脏差异表达序列标签

Identifying ESTs Differential Expressed in Liver Organ of Black-Bone Silky Fowl by Using Differential Displayed PCR

为了找出可能与丝羽乌骨鸡独特性状形成相关的基因,本研究运用差异显示PCR技术等对丝羽乌骨鸡与快大型肉鸡肝脏组织中差异表达的基因进行筛选,并通过生物信息学方法对所筛选出的差异表达序列标签进行分析。结果,本研究共筛选得到10条丝羽乌骨鸡与快大型肉鸡肝脏差异表达序列标签（dbEST登录号：CN606328~CN606329、CN606331~CN606338）。生物信息学分析结果显示：10条ESTs中,CN606332和CN606336是家鸡的18S rRNA基因部分序列,CN606333与编码家鸡热休克蛋白70的基因（HSPA5）同源,CN606328、CN606331与家鸡已有核酸数据库中的cDNA克隆或EST具有较高相似性,但功能未知。其他5条表达序列标签在家鸡核酸数据库中未发现相似序列,但分别与其他物种的细胞膜蛋白基因、γ-谷氨酰转肽酶（gamma-glutamyl transpeptidase）基因、ATP-binding cassette transporter基因、推定蛋白MGC27019基因以及异常纺锤型小脑畸形症（abnormalspindle microcephaly,ASPM）蛋白基因有不同程度的相似性。本研究结果表明,丝羽乌骨鸡与快大型肉鸡可能在多种代谢途径上都存在差异表达的基因,通过进一步深入研究这些差异表达基因,可以为研究丝羽乌骨鸡独特性状的形成提供重要的信息。

In order to find out genes related to special characters of the Black-Bone Silky Fowl,differential display PCR was used for screening differential expressed genes between liver organs of the Silkies and the AA broilers,and bioinformatics analysis were carried out to identify ESTs differential expressed.There were totally 10 ESTs（Acc_N：CN606328-CN606329,CN606331-CN606338）screened from the Silkies＇and the AA broilers＇liver organs.Results of bioinformatics analysis showed,among 10 ESTs,CN606332 and CN606336 were part of the chicken 18S rRNA gene;CN606333 was homologous with the chicken heat shock 70 ku protein 5（HSPA5）gene;CN606328 and CN606331 were significant similar with some of chicken cDNA clones or ESTs but had unknown function;the others had no significant similar with any chicken nucleotide sequences but had partly similarity to other species＇genes,including genes of membrane protein,gamma-glutamyl transpeptidase,ATP-binding cassette transporter,putative protein MGC27019,and abnormal spindle microcephaly protein（ASPM）.It was suggested in this study that many genes on varieties of metabolism pathways were differentially expressed between the Silkies and broilers.To study these genes more deeply could provide important information for studying the formation of special characters of the Silkies.