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异源二聚体蛋白白细胞介素-12在植物细胞悬浮培养中的表达与分泌

Expression and secretion of the heterodimeric protein interleukin-12 in plant cell suspension culture.

作者信息

Kwon T H, Seo J E, Kim J, Lee J H, Jang Y S, Yang M S

机构信息

Institute of Basic Sciences, Chonbuk National University, Jeonju 561-756, South Korea.

出版信息

Biotechnol Bioeng. 2003 Mar 30;81(7):870-5. doi: 10.1002/bit.10528.

Abstract

It has been suggested that plant cell culture is the most suitable system for producing small-to-medium quantities of specialized, expensive, and high-purity proteins. Here, we report that a heterodimeric protein, human interleukin-12 (hIL-12), was expressed and secreted into culture medium in a biologically active form. A transgenic plant expressing hIL-12 was constructed by sexual crossing of plants that expressed each subunit of the protein. From a piece of transgenic plant, callus was induced and cell suspension culture was established. The biological activity and amount of hIL-12 secreted into culture medium were analyzed using bioassays and ELISA. Analysis of cellular localization demonstrated that the protein was secreted into the culture medium together with its intrinsic signal peptide.

摘要

有人认为,植物细胞培养是生产中小批量特殊、昂贵且高纯度蛋白质的最合适系统。在此,我们报告一种异二聚体蛋白——人白细胞介素-12(hIL-12),以生物活性形式表达并分泌到培养基中。通过对表达该蛋白各亚基的植物进行有性杂交,构建了表达hIL-12的转基因植物。从一片转基因植物中诱导出愈伤组织,并建立了细胞悬浮培养体系。使用生物测定法和酶联免疫吸附测定法(ELISA)分析了分泌到培养基中的hIL-12的生物活性和含量。细胞定位分析表明,该蛋白与其内在信号肽一起分泌到了培养基中。

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