用实时荧光定量RT-PCR检测单个小鼠种植前胚胎中特异性基因表达  

Applying real-time fluorescence quantitative RT-PCR to test the specific gene expression of single mouse pre-implantation embryos

作　　者：陈林君[1] 孙海翔[1] 张宁媛[1] 徐志鹏[1] 

机构地区：[1]南京大学医学院附属鼓楼医院生殖医学中心,江苏南京210008

出　　处：《安徽医药》2010年第11期1328-1330,共3页Anhui Medical and Pharmaceutical Journal

摘　　要：目的用单细胞实时荧光定量RT-PCR,检测单个小鼠种植前胚胎中特异性基因表达。方法采集小鼠卵母细胞和原核期胚胎,后者经体外培养至囊胚。采用实时荧光定量RT-PCR对多个及单个卵母细胞和2-细胞期胚胎中Tcl1基因的表达进行定量检测;用同样方法对小鼠种植前各发育阶段的胚胎进行Tcl1基因的表达检测。结果在多个及单个卵母细胞和2-细胞期胚胎中,均检测出Tcl1的表达;多个卵母细胞、2-细胞期胚胎中Tcl1的表达量显著高于单个卵母细胞、2-细胞期胚胎;此方法也适用于胚胎早期发育的各个时期,Tcl1在单个卵母细胞、原核期胚胎和2-细胞期胚胎中表达量最高,在4-细胞期胚胎、8-细胞期胚胎、桑葚胚和囊胚中表达量极低。结论应用单细胞实时荧光定量RT-PCR可对单个小鼠种植前胚胎中基因的表达进行定量检测。Aim To test the specific gene expression of single mouse pre-implantation embryos by applying real-time fluorescence quantitative RT-PCR（rQ-RT-PCR）.Methods The mouse oocytes and pronuclear-stage embryos（zygote） which are cultured in vitro to blastocysts were collected.Then,the rQ-RT-PCR was used to test quantitatively the expression of gene Tcl1 in the oocytes and 2-cell stage embryos.The same methods were applied in the examination of gene Tcl1 in mouse embyos in every stage before implantation.Results Tcll was found quantitatively in multiple oocytes,single oocyte and 2-cell stage embryos.The expression of Tcll in multiple oocytes and 2-cell embryos was much higher than that in single oocyte and 2-cell embryo.The method could be applied in every embryo stage.The highest expression of Tcll was found in oocyte,pronuclear stage embryos and 2-cell stage embryos,however,its expression in 4-cell and 8-cell embryos,morulas and blastocysts was extremely low.Conclusion The rQ-RT-PCR can be applied to quantitative examination of gene expression in single mouse pre-implantation embryo.

关 键 词：实时定量 RT-PCR 小鼠 胚胎 Tcl1 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]