Ohuchi S J, Ikawa Y, Shiraishi H, Inoue T
Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8224, Japan.
Nucleic Acids Res Suppl. 2001(1):125-6. doi: 10.1093/nass/1.1.125.
Catalysis of Group I intron ribozymes is carried out by its core region consisting of two helical domains P4-P6 and P3-P7. Recently, our laboratory showed that a mutant Group I ribozyme lacking both the P4-P6 domain and the base-triples can perform the trans-esterification reactions. The result demonstrates that the elements required for splicing are concentrated in the P3-P7 domain. Based on this result, we carried out in vitro selection experiment starting from newly constructed libraries in the ribozyme lacking the P4-P6 domain and the base-triples. This selection experiment showed unexpected divergency on the limited sequence space for the active ribozymes.
I 组内含子核酶的催化作用由其核心区域完成,该核心区域由两个螺旋结构域P4 - P6和P3 - P7组成。最近,我们实验室表明,一种同时缺失P4 - P6结构域和碱基三联体的I组核酶突变体能够进行转酯反应。结果表明,剪接所需的元件集中在P3 - P7结构域。基于这一结果,我们从新构建的缺乏P4 - P6结构域和碱基三联体的核酶文库开始进行体外筛选实验。该筛选实验表明,活性核酶在有限的序列空间上出现了意想不到的差异。