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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (5): 19-22.

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

人β-防御素3基因的克隆分析及其真核表达载体的构建

黄仙仙 徐婷婷 徐雪丽 赵卫东 郑振宇 李春丽   

  • 收稿日期:2011-07-18 修回日期:2011-08-08 出版日期:2012-02-15 发布日期:2012-02-15
  • 基金资助:

    一种新型抗生素的研究—人防御素的克隆表达

Cloning and Sequence Analysis of Human β-defensin-3 Gene and Construction Its Eukaryotic Expression Vector

chunli li   

  • Received:2011-07-18 Revised:2011-08-08 Online:2012-02-15 Published:2012-02-15

PDF (PC)

796

摘要：

根据Genebank上的公布的人β防御素3基因序列设计引物，以提取的人DNA为模板进行PCR扩增，得到全长为1391bp的人β防御素3完整基因组序列，包括起始序列、信号肽序列，两个外显子、一个内含子以及上下游酶切位点和终止密码子等。与NCBI上公布的DNA序列的同源性达到99.93%，在内含子上有一个A的缺失，其编码氨基酸序列与公布序列的同源性达100%。将其克隆到pMD18-T载体中，进一步构建了其真核表达载体pcDNA-hBD3，为其表达和功能研究奠定基础。

关键词: 猪, 猪, 血源树突状细胞, 诱导, 鉴定

Abstract:

One pair of primer was synthesized according to the sequence of hBD3 published in Genebank. The 1391bp of human β-defensin-3 genomics sequence was obtained including the signal sequence, two exton ,one intron and restriction enzyme site and end code. Compared with the sequence published in NCBI, 99.93% nucleic acids were the same except for A deletion in intron. 100% amino acid were same. The sequence was first inserted into pMD18-T vector, then inserted into the pcDNA3.1 vector which resulted in the eukaryotic expression vector pCDNA-hBD3. This was the base for further study of its expression and function.