小鼠ERβ基因片段的克隆及其在胚胎发育中的表达  

Cloning of gene fragment of estrogen receptor-β and its expression in mouse embryo

作　　者：张子峰[1] 樊少华[1] 陆军[2] 吴冬梅[2] 单群[1] 胡斌[1] 李飞[1] 郑元林[1] 

机构地区：[1]徐州师范大学生命科学学院,徐州221116 [2]江苏省药用植物生物技术重点实验室

出　　处：《遗传》2008年第3期347-351,共5页Hereditas（Beijing)

基　　金：江苏省属高校自然科学重大基础研究项目(编号:2007KJA36029);江苏省青蓝工程中青年学术带头人培养项目(编号:2004);江苏省药用植物生物技术重点实验室课题;徐州师范大学自然科学基金重点项目(编号:07XLA09)资助~~

摘　　要：为探讨ERβ在小鼠胚胎发育过程中的表达,首先设计ERβ引物并扩增ERβ基因片段,构建ERβ/pGEM-3Z重组质粒进行克隆,分别用EcoRⅠ和HindⅢ进行酶切得到线性化DNA片段,以Sp6和T7聚合酶合成地高辛标记的(dig)正、反义RNA探针。然后通过胚胎整体原位杂交技术分析ERβ在小鼠胚胎中的表达。运用该探针检测到ERβ基因在10.5dpc胚胎的脑、脊神经管、生殖脊、心包、肢芽及颌弓部位表达,在13.5dpc胚胎的端脑、中脑、延髓、脊髓、肢芽中表达。推测ERβ基因可能在小鼠胚胎性别分化过程中起调节作用;可能在小鼠胚胎神经管的早期区域化过程中起作用并在3个原始脑泡进一步分化及脊髓的分化过程中起作用;可能在小鼠胚胎肢芽中的骨与软骨的形成与分化中起调控作用;可能在小鼠胚胎心脏发育过程中起作用,可能在小鼠胚胎颌弓的表面分化过程中起调控作用。In order to study the expression and regulation effects of estrogen receptor-β （ERβ） in the development of mouse embryo, the primer of ERfl was designed, the ERfl fragment was first obtained by RT-PCR and subcloned into plasmids pGEM- 3Z, then the recombinant plasmids were linearized with the restriction enzymes of EcoR Ⅰ and HindⅢ. Using Sp6 and T7 RNA polymerase, the digoxigenin（dig） labeled sense and anti-sense probes were transcriped in vitro, respectively. Then the expression of ERβ in mouse embryo was examined with the probes by whole-mount in situ hybridization. The results indicated that ERβ is expressed in the brain, spinal neural tube, genital ridge, pericardium, limb bud and mandibular arch of 10.5 dpc embryo, and is also expressed in the telencephalon, mesencephalon, medulla oblongata, spinal cord and limb bud of 13.5 dpc embryo. These results suggest that ERβ maybe play a role of regulation in sexual differentiation, primal differentiation of neural tube, further differentiation of three primary cerebral vesicles and spinal cord, generation and differentiation of bone and cartilage of limb bud, development of pericardium and configuration differentiation of mandibular in mouse embryo.

关 键 词：雌激素受体Β 基因克隆 整体原位杂交 小鼠胚胎 

分 类 号：Q78[生物学—分子生物学]