INVITRO SELF-SPLICING REACTIONS OF THE CHLOROPLAST GROUP-I INTRON CR LSU FROM CHLAMYDOMONAS-REINHARDTII AND INVIVO MANIPULATION VIA GENE-REPLACEMENT

被引：33

作者：

THOMPSON, AJ ^{[1
]}

HERRIN, DL ^{[1
]}

机构：

[1] UNIV TEXAS, DEPT BOT, AUSTIN, TX 78713 USA

来源：

NUCLEIC ACIDS RESEARCH | 1991年 / 19卷 / 23期

基金：

美国国家科学基金会;

关键词：

D O I：

10.1093/nar/19.23.6611

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The group I intron from the chloroplast rRNA large subunit of Chlamydomonas reinhardtii (Cr.LSU) undergoes autocatalytic splicing in vitro. Cr.LSU displays a range of reactions typical of other group I introns. Under optimal conditions, the 5' cleavage step proceeds rapidly, but the exon-ligation step is relatively slow, and no pH dependent hydrolysis of the 3' splice site occurs. A requirement for high temperature and high [Mg2+] suggests involvement of additional splicing factors in vivo. The positions of three cyclization sites of the free intron have been mapped; two of these sites represent reactions analogous to 5'-splice site cleavage, whereas the third is an example of G-exchange. Cr.LSU contains an open reading frame (ORF) potentially encoding an 163 amino acid polypeptide. ORF function has been investigated by using chloroplast gene replacement via particle bombardment. We have shown that the ORF can be deleted from Cr.LSU without affecting splicing in vivo and it thus does not encode an essential splicing factor.

引用

页码：6611 / 6618

页数：8

共 50 条

[1] THE CONSERVED U.G PAIR IN THE 5' SPLICE SITE DUPLEX OF A GROUP-I INTRON IS REQUIRED IN THE 1ST BUT NOT THE 2ND STEP OF SELF-SPLICING [J].