LINE1-ORF1抗体的制备及神经祖细胞分化中LINE-1转座水平的检测

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (8): 1083-1088.

LINE1-ORF1抗体的制备及神经祖细胞分化中LINE-1转座水平的检测

张海燕¹,陆菁菁²

1. 首都医科大学丰台教学医院
2. 首都医科大学附属北京天坛医院

收稿日期:2014-11-04 修回日期:2015-03-23 出版日期:2015-08-05 发布日期:2015-07-15
通讯作者: 陆菁菁 E-mail:lujingjing666@live.cn

Preparation of LINE1-ORF1 Antibody and detection of LINE-1 retrotransposition during neural progentitor cells differentiation

¹,Jingjing Lu

Received:2014-11-04 Revised:2015-03-23 Online:2015-08-05 Published:2015-07-15
Contact: Jingjing Lu E-mail:lujingjing666@live.cn

摘要/Abstract

摘要： 目的研究转座子LINE-1在神经细胞分化中的转座水平的变化。方法用原核表达系统表达LINE-1 ORF1蛋白，免疫后制备LINE-1 ORF1蛋白抗体，用Western blot法检测胚胎干细胞分化至神经祖细胞过程中LINE-1转座水平。结果制备的抗体具有较好的特异性和灵敏度，不同类型真核细胞中LINE-1转座水平不完全一致。同时，对细胞分化过程中的LINE-1 ORF1蛋白水平直接进行分析，首次在蛋白水平证实了神经细胞分化过程中LINE-1转座水平在分化前期增高，后期降低。结论 LINE-1在神经系统发育中具有较高的转座活性，提示，其在神经系统发育中扮演着重要的作用。

关键词: LINE-1, 转座, 神经祖细胞

Abstract: Objective To explore endogenous LINE-1 expression during Neural progenitor cells (NPCs) differentiation. Method recombinant LINE-1 ORF1 protein was expressed in bacteria, followed by purification, and the purified protein was used to immunize rabbits to produce anti-ORF1 polyclonal antibody. The antibody was used to detect LINE-1 retrotransposition during neural progenitor cells (NPCs) differentiation by Western blotting. Result The anti-ORF1 antibody possessed high sensitivity and specificity, and observed the variability of LINE-1 retrotransposition in different eukaryotic cell lines. Using anti-ORF1 antibody to detect LINE-1 ORF1 protein level during NPC differentiation, it was found that when stem cells commits to NPCs, LINE-1 retrotransiposition increases in the early stage and decline in the late stage. Conclusion LINE-1 retrotranspose increases upon neuronal commitment, suggested LINE-1 play a pivotal role in neural development.

Key words: LINE-1, retrotranspostion, neural progenitor cells

中图分类号:

R741

张海燕陆菁菁. LINE1-ORF1抗体的制备及神经祖细胞分化中LINE-1转座水平的检测[J]. 基础医学与临床, 2015, 35(8): 1083-1088.

Jingjing Lu. Preparation of LINE1-ORF1 Antibody and detection of LINE-1 retrotransposition during neural progentitor cells differentiation[J]. Basic & Clinical Medicine, 2015, 35(8): 1083-1088.