【摘要】 采用抑制消减杂交技术(suppression subtractive hybridization,SSH),以SD大鼠为实验材料,选取妊娠过程中植入前期(第5天,D5) 和分娩前期(第19天,D19) 子宫分别作为驱动方(driver) 和实验方(tester),进行抑制消减杂交,获得的消减文库经差异筛选得到70个阳性克隆. 序列测定和同源对比分析表明,这些克隆所代表的基因在大鼠基因库中分别与8个已知基因有90%～100%不等的同源性. 这些基因均差别表达于分娩前期子宫组织中,其中首次发现尿鸟苷蛋白和干扰素诱导蛋白16在SD大鼠妊娠子宫中有表达. RT-PCR及半定量分析显示,尿鸟苷蛋白基因在妊娠第19天子宫中的表达显著高于妊娠第5天(P<0.001),而干扰素诱导蛋白16差异表达不明显. 在妊娠D6、D9和D12的子宫中尿鸟苷蛋白基因自胚泡植入后其表达逐渐上升,妊娠D15下降,在妊娠D19表达量最高. 结果提示特异表达的尿鸟苷蛋白基因可能与分娩有关.更多还原

【Abstract】 SD rats were applied as experiment materials to screen differentially expressed genes of the uterus in different stage of pregnancy by suppression subtractive hybridization (SSH), in which the uterus from the 5th and the 19th day in pregnancy were respectively used as driver and tester. Seventy positive clones from forward subtractive library are selected by differential screening. Sequencing analysis and homology comparison showed that all these clones are homologous to eight known genes in GenBank by the degree from 90% to 100%. These genes are differentially expressed in the uterus from the 19th day in pregnancy, in which the expression of uroguanylin and interferon-inducible protein 16 in the uterus in pregnancy are reported for the first time. Semi-quantitative analysis showed that the expression of uroguanylin was significantly higher in the uterus of 19th day in pregnancy than that of 5th day (P < 0.001), however, there was no difference about the expression of interferon-inducible protein 16. The expression of uroguanylin increased after embryo implantation, decreased on the 15th day and reached the peak on the 19th day in pregnancy. The result suggested that uroguanylin might be concerned with parturition.更多还原