为研究通过抑制消减杂交筛选出的cdk2ap1基因在鸡胚胎发育过程中的表达, 设计cdk2ap1引物并通过 RT-PCR扩增cdk2ap1基因片段, 构建cdk2ap1/pGEM-T重组质粒。以构建的重组质粒为模板, 使用Sp6和T7 RNA聚合酶合成地高辛(dig )标记的正、反义RNA探针, 借助胚胎整体原位杂交技术研究了cdk2ap1在雌雄鸡胚中的表达。结果表明cdk2ap1基因在4.0 d两性胚胎的脑间质、菱脑、听囊、脊神经管、前肢等部位均有表达, 且在雄性鸡胚中的表达量高于雌性; 在雄性鸡胚的生殖脊、后肢部位中该基因有表达但在雌性胚胎对应部位却基本没有表达。鉴于该基因在雌雄鸡胚中的差异表达, 推测cdk2ap1基因在鸡胚性别分化及性腺发育过程中起一定调节作用。

To investigate the expression and functions of cyclin-dependent kinase 2-associated protein 1 (cdk2ap1) screened by suppression subtractive hybridization in chicken embryo development, a pair of primers was designed to amplify the cdk2ap1 fragment by RT-PCR and subsequently the fragment obtained was cloned into the plasmid pGEM-T. Sense and antisense probes labeled with digoxigenin were generated using SP6 and T7 RNA polymerases, respectively, and used to examine cdk2ap1 expression in chicken embryos of both sexes by whole-mount in situ hybridization. In both sexes, cdk2ap1 was expressed in the head mesenchyme, rhombencephalon, optic vesicles, spinal neural tube, and forelimb of 4.0-day-old embryos and the expression in males was significantly higher than that in females. In addition, in the genital ridge and hindlimb of the 4.0-day-old chicken embryo, cdk2ap1 was obviously expressed in the males but not in females. It is supposed that cdk2ap1 may play a role in the sexual differentiation and development of gonad of chicken embryo.