[1]张彦威,刘丽雪,赵琳,等.大豆GmGBP1基因的原核表达及重组蛋白纯化[J].大豆科学,2012,31(06):874-877.[doi:10.3969/j.issn.1000-9841.2012.06.003]
　ZHANG Yan-wei,LIU Li-xue,ZHAO Lin,et al.Prokaryotic Expression and Protein Purification of Soybean GmGBP1 Gene[J].Soybean Science,2012,31(06):874-877.[doi:10.3969/j.issn.1000-9841.2012.06.003]
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大豆GmGBP1基因的原核表达及重组蛋白纯化

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年06期 页码: 874-877 栏目: 出版日期: 2012-12-30
Title:
Prokaryotic Expression and Protein Purification of Soybean GmGBP1 Gene
文章编号:
1000-841（2012）06-874-4
作者:
张彦威; 刘丽雪; 赵琳; 谷月娇; 卢清瑶; 宋仙萍; 李文滨
东北农业大学大豆生物学教育部重点实验室，东北农业大学农业部大豆生物学与遗传育种重点实验室，黑龙江 哈尔滨150030
Author(s):
ZHANG Yan-wei; LIU Li-xue; ZHAO Lin; GU Yue-jiao; LU Qing-yao; SONG Xian-ping; LI Wen-bin
Key Laboratory of Soybean Biology in Chinese Ministry of Education,Key Laboratory of Soybean Biology and Breeding/Genetics of Chinese Agriculture Ministry,Northeast Agricultural University,Harbin 150030,Heilongjiang,China
关键词:
大豆; SKIP; 原核表达; 包涵体; 纯化
Keywords:
Soybean; SKIP; Prokaryotic expression; Inclusion body; Purification
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.06.003
文献标志码:
A
摘要:
将大豆中已克隆的一个新的SKIP基因(GmGBP1)构建到原核表达载体pGEX-6p-1上，导入大肠杆菌BL21(DE3)中,对其进行IPTG诱导。结果表明:在IPTG浓度为0.1 mmol.L-1,诱导时间为8 h，诱导温度为37℃时，重组蛋白得到表达，分子量大约为95 kDa，SDSPAGE电泳结果表明重组蛋白主要以包涵体形式存在，用8 mol.L-1尿素对其进行溶解后经GST柱纯化，得到较好的纯化效果。
Abstract:
A new SKIP homolog in soybean was constructed into prokaryotic expression vector pGEX-6p-1 and then transformed into E.coli BL21(DE3)to get the expression protein for further study.The result indicated that an 95 kDa recombinant protein was expressed with the treatment of 0.1 mmol.L-1IPTG for 8 h at 37℃,the recombinant protein was confirmed to mainly existed in inclusion body form by SDSPAGE.The high uality recombinant protein was obtained through GST column purification after the inclusion body was dissolved with 8 mol.L-1 urea.

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备注/Memo

收稿日期：2012-9-9
基金项目：国家自然科学基金（31101169, 30671318，31271748）；转基因生物新品种培育重大专项（2011ZX08004-005）；黑龙江省教育厅大豆分子设计团队项目；黑龙江省教育厅省高校青年学术骨干项目；东北农业大学博士基金。
第一作者简介：张彦威（1987-），男，在读博士，研究方向为大豆生物技术。E-mail：zywei1987@163.com。
通讯作者：李文滨(1958-)，男，教授，博士生导师，从事大豆遗传育种研究。E-mail：wenbinli@yahoo.com。

更新日期/Last Update: 2014-08-07