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《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:

第45卷

期数:

2017年08期

页码:

33-36

栏目:

生物技术

出版日期:

2017-04-22

文章信息/Info

Title:

Prokaryotic expression of Arabidopsis protein ERA-1 and preparation of its polyclonal antibodies

作者:

付世英;  刘夏燕

西北农林科技大学生命科学学院，陕西杨凌 712100

Author(s):

Fu Shiying; et al

关键词:

拟南芥ERA-1; 原核表达; 蛋白纯化; 抗血清; 多克隆抗体制备

Keywords:

-

分类号:

Q946.1

DOI:

-

文献标志码:

A

摘要:

将拟南芥ERA-1基因除信号肽外的剩余编码序列克隆到原核表达载体pET28a上，转化到大肠杆菌BL21 (DE3)中表达，发现表达的目的蛋白位于包涵体中，大量表达目的蛋白并用镍柱亲和层析法进行纯化，获得足量的重组蛋白后将其作为抗原免疫家兔，获得抗血清。用重组蛋白亲和纯化抗血清中的ERA-1多克隆抗体。利用野生型拟南芥、ERA-1敲除突变系和ERA-1过表达系的叶片总蛋白进行免疫印迹试验，检测ERA-1多克隆抗体的特异性，发现纯化后的ERA-1多克隆抗体(1 ∶[KG-*3]1 000体积比稀释)能特异地检测到拟南芥中的ERA-1蛋白。本试验成功制备了ERA-1蛋白的多克隆抗体，可以用于今后对ERA-1基因功能的研究。

Abstract:

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参考文献/References:

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备注/Memo

备注/Memo:

收稿日期：2016-03-21
作者简介：付世英(1989—)，女，河南修武人，硕士研究生，从事植物分子生物学研究。E-mail：shiyingfu123@163.com。
通信作者：刘夏燕，副教授，从事植物分子生物学研究。E-mail：xiayanliu@gmail.com。

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更新日期/Last Update: 2017-04-20