生物技术通报 ›› 2013, Vol. 0 ›› Issue (3): 186-191.

• 研究报告 • 上一篇    下一篇

肿瘤转移抑制基因KISS-1和NM23-H1的原核表达与纯化

左芳雷,冯秀娟,陈尚武   

  1. 中国农业大学食品科学与营养工程学院,北京 100083
  • 收稿日期:2012-09-20 修回日期:2013-03-21 出版日期:2013-03-20 发布日期:2013-03-21
  • 作者简介:左芳雷,男,博士研究生,研究方向:食品生物技术;E-mail :duruolanzi@gmail.com
  • 基金资助:
    国家自然科学基金资助项目(31071507)

Prokaryotic Expression and Purification of Metastasis Suppressor Gene KISS-1 and NM23-H1

Zuo Fanglei, Feng Xiujuan, Chen Shangwu   

  1. College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083)
  • Received:2012-09-20 Revised:2013-03-21 Published:2013-03-20 Online:2013-03-21

摘要: 从人胎盘组织和乳腺癌细胞中克隆得到肿瘤转移抑制基因KISS-1 和NM23-H1,经Blast 比对分析与GenBank 公布 序列同源性达100%。NM23-H1 在大肠杆菌中成功表达,大部分为可溶性蛋白,占总蛋白50% 以上,经纯化后纯度达到90% ;而 KISS-1 需要与载体的部分序列融合才能够得到有效表达,表达产物为不可溶的包涵体蛋白,占包涵体总量的70% 以上,经纯化后 KISS-1 蛋白可能形成多聚体结构。

关键词: 肿瘤转移抑制基因, KISS-1, NM23-H1, 表达, 纯化

Abstract: Metastasis suppressor gene KISS-1 and NM23-H1 were cloned from human placenta and breast cancer cell,share 100% identity with that published in GenBank. NM23-H1 was successfully expressed in E.coli, the major amount of protein was soluble, accounting for more than 50% of total protein, the purified protein has 90% purity ;however, KISS-1 should be fused to the sequence from expression vector then can effectively expressed, the expressed protein was insoluble inclusion body, accounting for more than 70% of total insoluble protein, after purification, KISS-1 may formed into polymeric structures.

Key words: Metastasis suppressor gene, KISS-1, NM23-H1, Expression, Purification