INVITRO CLEAVAGE OF HPV16 E6 AND E7 RNA FRAGMENTS BY SYNTHETIC RIBOZYMES AND TRANSCRIBED RIBOZYMES FROM RNA-TRIMMING PLASMIDS
INVITRO CLEAVAGE OF HPV16 E6 AND E7 RNA FRAGMENTS BY SYNTHETIC RIBOZYMES AND TRANSCRIBED RIBOZYMES FROM RNA-TRIMMING PLASMIDS
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DOI：
10.1016/0014-5793(93)81102-6
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发表时间：
1993-05-03
期刊：
影响因子：
3.5
通讯作者：
QI, GR
中科院分区：
文献类型：
--
作者：
HE, YK;LU, CD;QI, GR
构建了一种RNA修剪质粒pRG523，其中三个核酶基因GR5（5′ - 顺式核酶基因）、HR2G（反式核酶基因）和GR3（3′ - 顺式核酶基因）按从5′到3′的顺序排列在T7启动子下游。该质粒的体外转录表明，反式核酶可被其两侧的顺式核酶修剪至确定的长度。研究了合成核酶对不同长度的人乳头瘤病毒16型（HPV16）E6和E7 RNA片段的体外切割，以及合成核酶和具有不同长度侧翼序列的转录核酶对长度为171个核苷酸的E7 RNA的切割。结果表明，核酶和靶RNA上的非碱基配对侧翼序列均可影响切割反应。
A RNA-trimming plasmid pRG523 is constructed, in which three Rz genes, GR5(5'-cis-Rz gene), HR2G(trans-Rz gene) and GR3(3'-cis-Rz gene), are arranged in the order from 5' to 3' downstream from the T7 promoter. In vitro transcription of this plasmid shows that the trans-Rz can be trimmed to definite lengths by the cis-Rz on both sides of the trans-Rz. In vitro cleavage of HPV16 E6 and E7 RNA fragments of different lengths by synthetic Rz and that of E7 RNA with a length of 171 nt by synthetic Rz and transcribed Rzs with different lengths of flanking sequences is studied. The results show that the non-base-pairing flanking sequences on both Rz and target RNA can affect the cleavage reaction.