Ehrich E, Craig A, Poustka A, Frischauf A M, Lehrach H
Imperial Cancer Research Fund, London, U.K.
Gene. 1987;57(2-3):229-37. doi: 10.1016/0378-1119(87)90126-0.
A family of cosmid vectors was constructed which contain replication origins (ori) derived from the multicopy plasmid R6K, a kanamycin resistance gene and two cos sites, permitting efficient library construction. Additional features of later constructs are (i) the presence of NotI sites flanking the site of insertion to allow intact excision of inserts, (ii) the facility for selective cloning of the ends of inserts for rapid chromosome walking, and (iii) the use of a mutated R6K ori leading to an increased copy number.
构建了一个黏粒载体家族,其包含源自多拷贝质粒R6K的复制起点(ori)、一个卡那霉素抗性基因和两个黏性末端位点,从而实现高效文库构建。后续构建体的其他特征包括:(i)在插入位点两侧存在NotI位点,以便完整切除插入片段;(ii)具备选择性克隆插入片段末端以进行快速染色体步移的功能;(iii)使用了一个导致拷贝数增加的突变型R6K ori。