多拷贝策略在小肽表达中的应用.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2011, May 25; 27(5): 684−689 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn ©2011 CJB, All rights reserved. 综 述 多拷贝策略在小肽表达中的应用 曹艳萍，单安山，马清泉，尹佳佳 东北农业大学动物营养研究所，哈尔滨 150030 摘 要: 基因工程技术已经在大分子多肽表达上得到了广泛的应用。但是小分子多肽不稳定且易降解，使其表达后很 难检测和纯化。多拷贝策略是将目的基因或是含有目的基因的表达盒首尾串联，而串联构建多拷贝表达载体是目前解 决小分子多肽表达量少的有效方法。总结和比较非对称粘性末端互补法、接头连接法、同尾酶法和表达盒串联法在多 肽表达方面的应用情况，为小分子多肽的体外表达提供方法和思路。 关键词: 多拷贝，串联，多肽表达，应用 Application of multi-copies in expression of smaller peptides: a review Yanping Cao, Anshan Shan, Qingquan Ma, and Jiajia Yin Institute of Animal Nutrition , Northeast Agricultural University, Harbin 150030, China Abstract: The technology of genetic engineering has been widely used to express macromolecules such as enzymes. However, it is difficult to detect and purify the micromolecules such as small peptides, because of their instability and degradability. Construction of multi-copy recombinant expression plasmid can be achieved by inserting multiple target genes or expression cassette containing target genes with the same orientation into expression vector. This is effective to increase the expression level of small peptides. In this article we described four methods in order to provide some optional methods and ideas for the expression of active small peptides. Keywords: multi-copies, in series, expression of polypeptides, application 近年来，随着基因工程技术的发展，一些有生 保健、生物制药和临床应用等方面有着广阔的前景。 物活性的小分子多肽受到了高度的重视，国内外针 但是由于其分子量小，在细胞中的稳定性不好，容易 对小分子多肽的大量研究表明某些小肽在动物营养 被宿主蛋白酶识别并降解为无活性的寡肽片段[1] ，另 Received: October 23, 2010; Accepted: January 5, 2011 Supported by: National Natural Science Fou