为进一步研究干扰素α应答基因IFRG15(Interferon responsive gene 15)在小鼠整个发育过程中的表达规律,从植入前胚胎及2、5、16周龄的雌、雄昆明小白鼠心、肝、脾、肺、肾、肌肉、卵巢或睾丸等组织中提取总RNA,以HPRT1(Hypoxanthine phosphoribosyltransferase 1)为内参基因,利用RT-PCR的方法进行目的片段的扩增及差异性分析。结果表明,IFRG15在植入前胚胎8-细胞期,桑葚胚期开始显著高表达于受精卵、2-细胞期、4-细胞期(p〈0.05),在囊胚期表达量达到最高,且显著高于其他各期(p〈0.05);在雌雄小鼠几个组织体外发育过程中均检测到表达,但表达量有所不同,在雄性小鼠各组织中的表达无显著规律性差异;在5周龄雌性小鼠组织中达到最高(p〈0.05),卵巢组织尤为明显,推测该基因对卵巢的成熟有重要的促进作用;本实验成功获得IFRG15在小鼠植入前各期胚胎及体外发育过程中的表达模式,为进一步探究该基因在小鼠克隆胚发育过程中的作用奠定基础。
In order to study the expression pattern of IFRG15(Interferon responsive gene 15) during mouse development and illustrate the biological function of its encoded protein,RT-PCR method was used to detect the mRNA expression of IFRG15 in pre-implantation embryo and different mouse tissues with HPRT1(Hypoxanthine phosphoribosyltransferase 1)as internal control.Total RNA from pre-implantation embryo and heart,liver,spleen,lung,kidney,muscle,ovary or testis tissues of 2,5,16-week old female and male Kunming mice was extracted and used as templates for RT-PCR.The results showed that,in pre-implantation embryo,the expression of IFRG15 at 8-cell,morula stages was significantly higher than that in the zygote,2-cell,4-cell phases(p0.05);the expression in the blastocyst stage has become the highest,and significantly higher than that of the other stages(p0.05);meanwhile,IFRG15 was also found expressed in all the examined tissues,but the expression was different.In male mouse the expression had no significant differences in regularity.Besides,in 5-week old female mouse tissues the expression of the IFRG15 was found significantly higher than that in 2,16-week old mouse(P0.05),especially ovarian tissue.It suggested that IFRG15 has an important role in on ovarian maturation.This study successfully obtained the expression patterns of IFRG15 in mouse pre-implantation embryos and some tissues in vitro development.