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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (18): 194-198.doi: 10.11924/j.issn.1000-6850.2011-3143

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

植物甜蛋白马宾灵(MabinlinⅡ)多克隆抗体的制备与鉴定

顾文亮 夏启玉 姚晶 胡新文 郭建春   

  • 收稿日期:2011-10-27 修回日期:2011-12-14 出版日期:2012-06-25 发布日期:2012-06-25
  • 基金资助:

    马宾灵基因(Mabinlin)体外甜味表达及其结构发生规律研究;分子伴侣SUMO融合表达马宾灵(MabinlinⅡ)的研究

Preparation and Identification of the Polyclonal Antibody Against Plant Sweet Protein MabinlinⅡ

  • Received:2011-10-27 Revised:2011-12-14 Online:2012-06-25 Published:2012-06-25

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摘要：

马宾灵(MabinlinⅡ)是中国所特有的植物马槟榔种子中的甜味蛋白，将其作为新型甜味剂有着广阔的市场前景。为了给重组马宾灵的基因工程应用提供可靠的蛋白质检测与鉴定的抗体，通过离子交换法从马槟榔(Capparis masaikai Lévl)种子中分离纯化马宾灵，将其作为抗原免疫新西兰大白兔，收集免疫后血清经抗原亲和纯化制备马宾灵多克隆抗体。结果表明，制备的马宾灵多克隆抗体经ELISA检测效价比达到1:256000，Western-blot检测表明其具有良好的反应性和特异性。本研究制备的马宾灵多克隆抗体可用于马宾灵在不同生物反应器中表达的检测，为马宾灵的基因工程研究提供灵敏可靠的免疫学鉴定。

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Abstract:

The plant sweet protein MabinlinⅡ is found rich in mature seeds of Capparis masaikai Lévl which is particular to China and may have wide application prospects in food sweeteners industry. In this study, in order to provide a reliable testing antibody for engineering applications of recombinant MabinlinⅡ, MabinlinⅡ was separated and purified from the mature seeds of Capparis masaikai Lévl by ion exchange chromatography. The purified protein was used as antigen to immunize New Zealand rabbits to prepare polyclonal antibody against the MabinlinⅡ protein. The result of Enzyme-Linked Immunosorbnent Assay (ELISA) showed that the titer of the polyclonal antibody to purified MabinlinⅡ was 1:256000, and the polyclonal antibody against the purified MabinlinⅡ also had highly reactivity and specialty in western blot analysis. The polyclonal antibody against MabinlinⅡ could be used for detecting the protein amounts of MabinlinⅡ expressed in various bioreactors, which may serve sensitive and reliable methods for immunology detection in genetic engineering research of MabinlinⅡ.