位点特异性重组系统的机理和应用  

Site-Specific Recombination Systems: Mechanisms and Applications

作　　者：张霖[1] 赵国屏[1,2,3] 丁晓明[1] 

机构地区：[1]复旦大学生命科学学院微生物学与微生物工程学系,上海200433 [2]中国科学院上海生命科学研究院植物生理生态研究所合成生物学重点实验室,上海200032 [3]国家人类基因组南方研究中心省部共建疾病与健康基因组学重点实验室,上海201203

出　　处：《中国科学：生命科学》2010年第12期1090-1111,共22页Scientia Sinica(Vitae)

基　　金：国家高技术研究发展计划(批准号:2007AA021206);国家自然科学基金(批准号:30600009和30830002)资助项目

摘　　要：位点特异性重组酶识别特定的位点形成联会复合体,并发生DNA链的切割与交换,实现靶位点之间的整合、切离或倒位.这一过程由位于重组酶催化活性中心的酪氨酸或丝氨酸向DNA磷酸骨架发起攻击,形成共价中间体,不需要高能量辅助因子的参与.由于位点特异性重组系统具有高效精确的优点,在基因工程领域得到了广泛的应用.本文从识别位点的性质、重组酶的组成与结构及催化反应的特点三方面对位点特异性重组的机理进行了全面的阐述,并对当前重组酶的应用研究之现状、热点及存在的问题作了深入剖析,并讨论了未来的发展趋势.Site-specific recombinases mediate integration, excision and inversion of target sites by recognizing the defined attachment sites, forming synaptic complexes followed by DNA cleavage and rejoining. Tyrosine or serine within the active center of recombinases attacks the sugar-phosphate backbone to form covalently linked intermediates, without the accessory of high-energy cofactors. Site-specific recombination systems have been widely used in genetic engineering attributed to the high efficiency and specificity of the reaction. The mechanisms of site-specific recombination are described based on the properties of recognition sites, the compositions and structures of recombinases, and the features of catalytic processes. The current status, research hot spots and problems of developing site-specific recombination systems for applications are extensively reviewed, and the future trends are discussed.

关 键 词：位点特异性重组 酪氨酸重组酶 丝氨酸重组酶 整合酶 

分 类 号：Q75[生物学—分子生物学]