基于Red重组系统进行基因替换的方法建立

A Method for Establishment of Gene Replacement Based on Red Recombinant System

快速、高效替换目的基因是大肠杆菌基因工程菌研究的前提和基础。该试验利用Red重组系统结合基因工程技术替换基因工程菌苗中的氯霉素抗性基因,同时留下半乳糖筛选标记基因,结果在Red重组系统的帮助下成功实现了半乳糖筛选标记基因对氯霉素抗性基因的置换,同时菌苗的其他表型特征未发生任何改变。该方法的建立将为具有新遗传特性的工程菌株和基因功能研究提供有力的工具。

The rapid and highly-efficient gene replacing is prerequisite for studying E. coli gene engineering strains. The Red recombinant system was combined with genetic engineering techniques to substitute chloromycetin-resistant gene from genetic engineering strain and simulta- neously maintain galactosc-screening marker. We have successfully confirmed that chloromycetin-resistant gene could be replaced by emplo- ying Red recombinant system and simultaneously retained other characteristic of strain, thus providing important insights into genetic engineer- ing strain with new genetic characteristics and gene function research.