CAJ | 학술논문

由λ噬菌体的exo、beta、gam 3个基因分别编码的Exo、Beta和Gam蛋白组成λ Red同源重组系统。λRed重组是细菌靶基因置换的有效技术，该技术发展迅速，已广泛应用于大肠埃希菌等细菌染色体DN A的基因敲除、基因整合，可以对任何大小的DN A分子进行精确修饰。利用λ Red重组系统不仅可以构建工程菌，还可以为研究细菌的致病机理和耐药机制提供新的方法。然而，许多应用因重组频率而受到限制，影响重组频率的因素主要有底物浓度、同源片段的长度、内源性核酸酶、错配蛋白等。论文综述了λ Red同源重组技术的应用进展及影响重组的因素，以期为优化重组试验方案及提高实验的成功率提供帮助。
유λ서균체적exo、beta、gam 3개기인분별편마적Exo、Beta화Gam단백조성λ Red동원중조계통。λRed중조시세균파기인치환적유효기술，해기술발전신속，이엄범응용우대장애희균등세균염색체DN A적기인고제、기인정합，가이대임하대소적DN A분자진행정학수식。이용λ Red중조계통불부가이구건공정균，환가이위연구세균적치병궤리화내약궤제제공신적방법。연이，허다응용인중조빈솔이수도한제，영향중조빈솔적인소주요유저물농도、동원편단적장도、내원성핵산매、착배단백등。논문종술료λ Red동원중조기술적응용진전급영향중조적인소，이기위우화중조시험방안급제고실험적성공솔제공방조。
TheλRed homologous recombination system is consisted of Exo ,Beta and Gam protein ,which are encoded by exo ,beta ,gam in theλphage respectively .This system has got an increasing development because of its efficiency in gene replacement of bacteria .It has been widely used in bacterial gene replace‐ment including gene knockout and gene integration in E .coli genomic DNA ,precise modification without consideration of any molecule weight .TheλRed homologus recombination system can not only build engi‐neering bacteria ,but also can provide a new insight for mechanisms of drug resistance and pathogenicity in bacteria .However ,many applications are limited by its low recombination frequency .Reports indicated that many factors contribute to recombination such as concentration of substrate ,length of homologous fragment ,endogenous nucleases ,mismatch protein and so on .In this paper ,the application and factors , which may influenceλRed homologous recombination ,were discussed in detail ,and aimed to optimize pro‐tocol and improve the efficiency in experiments .