发明名称 |
In vitro assay for inhibitors of the intron self-splicing reaction in Pneumocystis carinii |
摘要 |
The present invention pertains to an in vitro method for assaying for an inhibitor of the catalytic Group I self-splicing intron reaction in the nuclear rRNA genes of Pheumocystis carinii which comprises the steps of (a) providing a DNA template containing the intron (I) from the 26S rRNA gene in Pneumocystis carinii and a portion of the 5' and 3' flanking exons (E1 and E2, respectively) between nucleotides 1963 and 2267 of 26S rRNA (660 nucleotides of amplified rRNA gene including the group I intron); (b) preparing an RNA precursor by transcription of the DNA template in the presence of labeled nucleoside triphosphates to produce a labeled RNA precursor (E1-I-E2); (c) purifying the RNA precursor; (d) incubating the RNA precursor and the inhibitor in the presence of guanosine triphosphate and magnesium ions; and (e) determining the degree of inhibition by the inhibitor on the intron splicing reaction in the RNA precursor by measuring the amount of labeled splicing intermediates and splicing products.
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申请公布号 |
US5849484(A) |
申请公布日期 |
1998.12.15 |
申请号 |
US19950491690 |
申请日期 |
1995.06.19 |
申请人 |
UNIVERSITY OF MEDICINE & DENTISTRY OF NJ |
发明人 |
LEIBOWITZ, MICHAEL J.;LIU, YONG |
分类号 |
C07K14/44;C12Q1/68;(IPC1-7):C12Q1/68;C12P19/34;C07H21/04 |
主分类号 |
C07K14/44 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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