PBDC1蛋白的表达及其多克隆抗体的制备

基础医学与临床 ›› 2014, Vol. 34 ›› Issue (2): 222-225.

PBDC1蛋白的表达及其多克隆抗体的制备

戚武林,胡江江,曹玲玲,赵辅昆,张世馥

浙江理工大学生命科学学院

收稿日期:2013-08-05 修回日期:2013-09-24 出版日期:2014-02-05 发布日期:2014-01-13
通讯作者: 张世馥 E-mail:cklzhang@163.com
基金资助:
浙江省生物医学开放基金

Expression of PBDC1 protein and preparation of its polyclonal antibody

Received:2013-08-05 Revised:2013-09-24 Online:2014-02-05 Published:2014-01-13

摘要/Abstract

摘要： 目的原核表达、纯化PBDC1蛋白，制备PBDC1多克隆抗体。方法将PBDC1基因克隆到pET-28a(+)质粒中，构建成重组质粒pET-28a(+)-PBDC1。将此重组质粒转化大肠杆菌BL21(DE3)后，IPTG诱导其在宿主菌中大量表达，并进行SDS-PAGE检测。经镍离子亲和柱纯化得到PBDC1融合蛋白，并以此免疫新西兰大白兔制备多克隆抗体。结果经质谱鉴定，获得了高纯度的PBDC1蛋白。经Western blot验证，获得了抗PBDC1蛋白的多克隆抗体。结论成功获得抗PBDC1蛋白的多克隆抗体，为研究PBDC1在红系分化中的功能提供了有利工具。

关键词: PBDC1蛋白, 多克隆抗体

Abstract: Objective To express and purify PBDC1 protein by prokaryotic expression system, and to prepare polyclonal antibody. Methods The gene coding sequence of PBDC1 was inserted into pET-28a(+) vector to generate the pET-28a(+)-PBDC1 recombinant plasmid. Then PBDC1 protein was expressed in E.coli BL21(DE3) by IPTG induction and detected by SDS-PAGE. The fusion protein was purified by Ni2+ affinity chromatography. New Zealand rabbits were immuized with PBDC1 protein and the antiserum was obtained. Results Highly purified PBDC1 protein was obtained and identified by mass-spectrum. Western blot verified that the polyclonal antibody can specifically recognize PBDC1 protein. Conclusions The polyclonal antibody against PBDC1 is successfully prepared, which provides an efficient reagent for futher study of its role in erythroid differentiation.

Key words: PBDC1 protein, polyclonal antibody

中图分类号:

Q511

戚武林胡江江曹玲玲赵辅昆张世馥. PBDC1蛋白的表达及其多克隆抗体的制备[J]. 基础医学与临床, 2014, 34(2): 222-225.

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