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[1]杨舸,李晶,吕林峰,等.播娘蒿DsCOR蛋白纯化、抗体制备及对冷诱导的响应[J].应用与环境生物学报,2010,16(03):337-341.[doi:10.3724/SP.J.1145.2010.00337]
 YANG Ge,LI Jing,LÜ,et al.Purification of DsCOR in Descurainia sophia, Preparation of Antibody and Its Response to Cold Inducement[J].Chinese Journal of Applied & Environmental Biology,2010,16(03):337-341.[doi:10.3724/SP.J.1145.2010.00337]
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播娘蒿DsCOR蛋白纯化、抗体制备及对冷诱导的响应()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
16卷
期数:
2010年03期
页码:
337-341
栏目:
胁迫抗性生理及分子机制专栏
出版日期:
2010-06-25

文章信息/Info

Title:
Purification of DsCOR in Descurainia sophia, Preparation of Antibody and Its Response to Cold Inducement
作者:
杨舸李晶吕林峰王乐宋玲玲乔代蓉曹毅
(四川大学生命科学学院微生物与代谢工程四川省重点实验室 成都 610064)
Author(s):
YANG Ge LI Jing LÜ Linfeng WANG Le SONG Lingling QIAO Dairong CAO Yi
(Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu 610064, China)
关键词:
播娘蒿DsCOR基因蛋白纯化多克隆抗体冷诱导抗寒性
Keywords:
Descurainia sophia DsCOR gene protein purification polyclonal antibody cold -inducement freezing tolerance
分类号:
Q949.748.305 : Q78
DOI:
10.3724/SP.J.1145.2010.00337
文献标志码:
A
摘要:
为揭示播娘蒿(Descurainia sophia)极强的抗寒特性,在大肠杆菌中重组表达已克隆的播娘蒿抗寒基因DsCOR,比较Ni亲合层析和煮沸–透析袋电洗脱蛋白纯化方法,制备纯化的DsCOR重组蛋白多克隆抗体,Western Blotting检测播娘蒿在寒冷胁迫下DsCOR的表达特性. 结果表明,两种纯化技术均能纯化DsCOR蛋白,透析袋电洗脱法纯化蛋白浓度为1.21 mg/mL,是Ni亲和层析法纯化蛋白浓度的2倍,且经PEG8000浓缩后的蛋白总量分别为9.075 mg和4.256 mg;制备的DsCOR抗体效价达1 : 12 800,Western Blotting检测显示只有经过低温冷诱导的播娘蒿DsCOR蛋白才表达,证明了DsCOR基因受低温诱导的规律. 图4 表1 参15
Abstract:
In order to reveal the extreme freezing tolerance of Descurainia sophia, Escherichia coli BL21-pET-32a-DsCOR was induced to express recombinant protein with IPTG by using our previously cloned gene DsCOR (cold-regulated gene) from D. sophia. Purification of recombinant protein DsCOR and preparation of DsCOR polyclonal antibody were conducted by comparing the methods of immobilized Ni2+ affinity chromatography and the boiling-electroelution in dialysis bag, and then the expression characteristics of this protein in D. sophia under cold stress were monitored by Western blotting. The results showed that the two methods employed were both able to purify protein DsCOR, but the concentration of the purified protein by boiling-electroelution in dialysis bag, 1.21 mg/mL, was twice as that by immobilized Ni2+ affinity chromatography. Moreover, the total amount of the purified protein by the latter method was merely 4.256 mg which was not enough to prepare the polyclonal antibody. The valence of antibody reached 1 : 12 800 and protein DsCOR in D. sophia could only express under cold inducement. Fig 4, Tab 1, Ref 15

参考文献/References:

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备注/Memo

备注/Memo:
国家自然科学基金项目(Nos. 30871321,30771312,30971817)和国家重点基础研究发展计划(973计划)(No. 2009CB125910)资助 Supported by the National Natural Science Foundation of China (Nos. 30871321, 30771312, 30971817), and the National Basic Research & Development Program of China (973 Program, No. 2009CB125910)
更新日期/Last Update: 2010-06-23