Robertson M P, Hesselberth J R, Ellington A D
Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA.
RNA. 2001 Apr;7(4):513-23. doi: 10.1017/s1355838201002199.
A small ribozyme ligase (L1) selected from a random sequence population appears to utilize non-Watson-Crick base pairs at its ligation junction. Mutational and selection analyses confirmed the presence of these base pairings. Randomization of the L1 core and selection of active ligases yielded highly active variants whose rates were on the order of 1 min(-1). Base-pairing covariations confirmed the general secondary structure of the ligase, and the most active ligases contained a novel pentuple sequence covariation. The optimized L1 ligases may be optimal within their sequence spaces, and minimal ligases that span less than 60 nt in length have been engineered based on these results.
从随机序列群体中筛选出的一种小型核酶连接酶（L1），在其连接位点似乎利用了非沃森-克里克碱基对。突变和筛选分析证实了这些碱基配对的存在。对L1核心区域进行随机化处理并筛选活性连接酶，得到了活性很高的变体，其反应速率约为1分钟-1 。碱基配对共变证实了连接酶的一般二级结构，最具活性的连接酶含有一个新的五重序列共变。优化后的L1连接酶在其序列空间内可能是最优的，基于这些结果已构建出长度小于60个核苷酸的最小连接酶。
