应用抑制性消减杂交技术克隆大鼠肝再生过程中特异表达基因

Cloning Differentially Expressed Genes by Suppression Subtractive Hybridization in Rat Liver Regeneration

应用抑制性消减杂交技术成功地构建了高消减效率的正向消减cDNA文库 ,从随机挑取的 5 0个克隆中有31个均检出了 6 0～ 40 0bp插入片段 ,对这些插入cDNA片段进行测序后经Genbank同源性检索 ,表明其中 7个片段为未知新序列。大鼠肝切除后肝再生cDNA正向消减文库的建立为进一步大批量筛选、克隆肝再生特异性表达的未知新基因奠定了基础 。

The cDNA from rat regenerating liver tissue was used as the tester and that from normal liver was used as the driver.A highly efficient subtractive cDNA library was constructed by suppression subtractive hybridization(SSH).After screening,31 clones from 50 clones which were derived from the cDNA library were inserted by 60～400bp cDNA fragments.24 cDNA fragments corresponded to known genes and 7 cDNA fragments were unknown sequences(GenBank accession number:BG447490～447496).