p21基因在野生型斑马鱼胚胎发育过程中的表达  

The Expression of p21 in Development of Wild Type Zebrafish Embryo

作　　者：宋锦[1,2,3] 姬牧远[2,4] 吴西军 周艳华[2] 舒莉萍[1,2,5] 何志旭[1,2,4] 

机构地区：[1]贵州医科大学免疫学教研室,贵州贵阳550004 [2]贵州医科大学干细胞与组织工程实验中心,贵州贵阳550004 [3]贵州医科大学 [4]贵州医科大学附院儿科学教研室,贵州贵阳550004 [5]贵州医科大学实验动物中心,贵州贵阳550004

出　　处：《贵阳医学院学报》2016年第7期765-769,共5页Journal of Guiyang Medical College

基　　金：国家自然科学基金项目(30960412);贵州省科技基础条件平台项目[(2009)4005];贵州省优秀科技教育人才省长专项资金(S2008-4)

摘　　要：目的:探究p21基因在斑马鱼早期发育过程中的表达。方法:Trizol法提取野生型斑马鱼胚胎总RNA,RT-PCR法扩增p21基因片段,与p^(CS2+)载体重组构建p21-p^(CS2+)重组质粒,以T3 RNA聚合酶制备地高辛标记的反义mRNA探针,通过斑马鱼全胚胎原位杂交检测p21基因在野生型斑马鱼胚胎早期发育过程中的表达。结果:成功构建了p21-p^(CS2+)重组质粒,并制备反义mRNA探针;原位杂交结果显示野生型斑马鱼0.5~18 hpf的胚胎均未发现p21基因阳性杂交信号,24~36 hpf胚胎可在脊索部位观察到阳性信号,48~72 hpf胚胎可在头部胸腺附近观察到阳性信号,表达量均较低。结论:成功制备了p21-p^(CS2+)重组质粒及斑马鱼p21基因反义mRNA探针,p21基因在野生型斑马鱼胚胎不同发育时相中表达水平不同。Objective：To explore the expression of p21 in zebrafish. Methods：Extract 0.75 hpf-120 hpf wild-type Zebrafish embryos in multiple phase and total RNA by Trizol,amplificating p21 gene by RT-PCR,and constructing p21-pCS^2＋lasmid. Preparing antisense mRNA probe with T3 RNA pol-ymerase,using zebrafish whole-mount in situ hybridization to examine the p21 expression pattern in early embryo development. Results：Successfully constructed p21-pCS^2＋lasmid,preparing antisense mRNA probe;within situ hybridization,there is no expression of p21 in wild-type zebrafish at 0. 5 hpf-18 hpf;positive signal in notochord was found in 24 hpf-36 hpf of the wild-type Zebrafish embry-os,and positive signal could be found near head and thymus,but indicating low expression. Conclu-sion：In this research,it has been successfully constructed p21-pCS^2＋plasmid,antisense mRNA probe preparation,and have a preliminary observation in the expression of p21 gene in Zebrafish em-bryos in different periods.

关 键 词：P21 斑马鱼 胚胎 发育 探针 质粒 

分 类 号：R394.3[医药卫生—医学遗传学]