小鼠<em>Dazl</em>基因真核表达载体的构建及表达检测

doi:10.7668/hbnxb.2014.S1.008

华北农学报 ›› 2014, Vol. 29 ›› Issue (S1): 35-39. doi: 10.7668/hbnxb.2014.S1.008

所属专题： 生物技术；

小鼠Dazl基因真核表达载体的构建及表达检测

孙星虹^1,2, 孙晓凤^1,3, 孙雷雷⁴, 李兰^1,2

1. 青岛农业大学山东省高校动物生殖与种质创新重点实验室, 山东青岛 266109;
2. 青岛农业大学动物科技学院, 山东青岛 266109;
3. 青岛农业大学生命科学学院, 山东青岛 266109;
4. 中国共产党潍坊市坊子区委党校, 山东潍坊 261200

收稿日期:2014-09-28 出版日期:2014-12-28
通讯作者: 李兰(1977-),女,陕西汉中人,实验师,博士,主要从事动物生殖生物学研究。
作者简介:孙星虹(1987-),女,山东威海人,硕士,主要从事动物生殖生物学研究。
基金资助:
山东省优秀中青年科学家科研奖励基金项目(BS2010NY010);国家自然科学基金青年科学基金项目(31101716;31001010)

Construction of Mouse Dazl Eukaryotic Expression Vector and Expression

SUN Xing-hong^1,2, SUN Xiao-feng^1,3, SUN Lei-lei⁴, LI Lan^1,2

1. Key Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, Qingdao Agricultural University, Qingdao 266109, China;
2. College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China;
3. College of Life Science, Qingdao Agricultural University, Qingdao 266109, China;
4. Fangzi District Party School of CPC, Weifang 261200, China

Received:2014-09-28 Published:2014-12-28

摘要/Abstract

摘要： 为研究小鼠Dazl基因的功能及探究Dazl基因过表达在干细胞向生殖细胞分化过程中的作用,采用RT-PCR方法,从13.5 dpc 雌性胎鼠卵巢中克隆出Dazl基因的全部编码区,测序正确后利用限制性内切酶将其连接到真核表达载体pcDNA3.1上。线性化后对小鼠成纤维细胞进行转染,qRT-PCR及Western Blot技术显示外源基因Dazl已经成功转染并表达。为研究Dazl基因的功能及干细胞向生殖细胞的诱导分化奠定了基础。

关键词: Dazl基因, 真核表达载体, 转染

Abstract: To study the function of Dazl gene and to confirm its promotion role during the differentiation of stem cells to germ cells,the full coding sequence of Dazl gene was cloned from the ovaries of female mice of 13.5 days post coitum (dpc).Then it was ligated into eukaryotic expression vector pcDNA3.1 after digested by the restriction endonuclease Hind Ⅲ and Kpn Ⅰ.The mouse fibroblasts were transfected using the linearized pcDNA3.1-Dazl.The results of qRT-PCR and Western Blot showed the foreign gene Dazl had transfected into eukaryotic cells.The study would provide the foundations for the induction and differentiation of stem cells to germ cells.

Key words: Dazl, Eukaryotic expression vector, Transfection

中图分类号:

孙星虹, 孙晓凤, 孙雷雷, 李兰. 小鼠Dazl基因真核表达载体的构建及表达检测[J]. 华北农学报, 2014, 29(S1): 35-39. doi: 10.7668/hbnxb.2014.S1.008.

SUN Xing-hong, SUN Xiao-feng, SUN Lei-lei, LI Lan. Construction of Mouse Dazl Eukaryotic Expression Vector and Expression[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(S1): 35-39. doi: 10.7668/hbnxb.2014.S1.008.

http://www.hbnxb.net/CN/Y2014/V29/IS1/35

参考文献

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