应用mRNA差异显示技术研究精子发生中阶段特异性基因表达

Application of mRNA differential display in study of stage-specific gene expression during spermatogenesis

为进行精子发生过程中具有阶段特异性表达特征的基因研究，本文应用mRNA差异显示与曲细精管透光分期技术相结合的方法，对大鼠IX～XII期和XIII～Ⅰ期曲细精管的基因表达指纹图谱进行了比较。通过27组不同引物组合的差异PCR反应，共得到56个具有阶段特异性表达特征的cDNA片段。通过对它们中的12个进行序列测定和序列同源性分析，其中8个为代表未知基因的新的EST，另有一个与小鼠quaking基因具有高度的同源性，提示这一基因同样存在于大鼠曲轴精管中并呈阶段特异性表达。将所得序列送至GenBank，均获得新的序列编号。

In attempting study of stage-specific gene expression during spermatogenesis, we spplied the mRNAdifferential display, combined transillumination-assisted microdissection of seminiferous tubules, to investigate thedifferential gene expression between the pool of seminiferous tubule segments from stage IX^XII and that from stageXIII~ I. Twenty-seven different primer combinations were assayed and fifty-six cDNA that expressed in stage-specificmanner were obtained. We partially sequenced 12 cDNA and compared the cDNAs to sequence database. Amongthem, eight cDNAs represented novel sequences having no significant sequence similarity to any sequence includewithin Gen Bank database, one was identical to the part sequences of mouse quaking gene, suggesting the quaking genewas also expressed in rat seminiferous tubule in stage-specific manner. All of them were submitted to GenBank andobtained new accession number.