生物技术通报 ›› 2015, Vol. 31 ›› Issue (8): 102-107.doi: 10.13560/j.cnki.biotech.bull.1985.2015.08.015

• 研究报告 • 上一篇    下一篇

鸡Vezf1基因RCAS干扰载体构建及鉴定

张超, 葛君, 袁明婧, 叶军, 袁立   

  1. 厦门大学生命科学学院 细胞应激生物学国家重点实验室,厦门 361102
  • 收稿日期:2014-11-25 出版日期:2015-08-21 发布日期:2015-08-22
  • 作者简介:张超,男,博士研究生,研究方向:血管分子发育生物学;E-mail:zhangchao_xmu@aliyun.com
  • 基金资助:

    国家自然科学基金项目(30671021),福建省自然科学基金项目(2007J0113)

Identification and Construction of RCAS RNA Interference Vector for Chicken Vezf1

Zhang Chao Ge Jun Yuan Mingjing Ye Jun Yuan Li   

  1. School of Life Sciences,State Key Laboratory of Cellular Stress Biology,Xiamen University,Xiamen 361102
  • Received:2014-11-25 Published:2015-08-21 Online:2015-08-22

摘要:

为研究Vezf1基因在鸡胚早期发育过程中的作用,构建了针对鸡Vezf1基因的RCAS病毒RNA干扰载体,分别在鸡胚细胞和胚胎水平对Vezf1基因实施沉默,通过Real-time PCR和原位杂交法检测目的基因mRNA表达水平。结果表明,基于RCAS病毒的干扰载体可以在鸡胚成纤维细胞和活体鸡胚中成功沉默Vezf1基因的表达。此研究为利用鸡胚模型深入了解Vezf1基因在早期发育过程中的作用提供了素材。

关键词: RCAS载体, RNAi, Vezf1, 鸡胚

Abstract:

In order to study the role of Vezf1 during the earlier development of chicken embryo, RNA interference vector for the chicken Vezf1 was constructed based on RCAS viral vector system, and the gene silencing of chicken Vezf1 was carried out in both embryonic cells and chicken embryos, and the mRNA expression was measured by Real-time PCR and in situ hybridization. Results showed that this interference vector from RCAS virus could keep Vezf1 expression silencing successfully in chicken embryo fibroblasts and chick embryos. This study provides a fundamental data for studying the Vezf1’s role during early embryonic development using the chicken model.

Key words: RCAS vector, RNAi, Vezf1, chicken embryo