遗传 ›› 2009, Vol. 31 ›› Issue (9): 936-940.doi: 10.3724/SP.J.1005.2009.00936

• 研究报告 • 上一篇    下一篇

Cdk2ap1在不同性别鸡胚中的表达

杨宇;俸艳萍;龚萍;黄潘;李世军;彭秀丽;龚炎长

  

  1. 华中农业大学动物遗传育种与繁殖教育部重点实验室, 武汉 430070

  • 收稿日期:2008-12-10 修回日期:2009-04-18 出版日期:2009-09-10 发布日期:2009-09-10
  • 通讯作者: 龚炎长

Expression of cyclin-dependent kinase 2-associated protein 1 in chicken embryos of different sexes

YANG Yu;FENG Yan-Ping;GONG Ping;HUANG Pan;LI Shi-Jun;PENG Xiu-Li; GONG Yan-Zhang   

  1. Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China
  • Received:2008-12-10 Revised:2009-04-18 Online:2009-09-10 Published:2009-09-10
  • Contact: GONG Yan-Zhang

摘要:

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。

关键词: 性腺发育, 整体原位杂交, 鸡胚, 基因克隆, cdk2ap1

Abstract:

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.